A post-MORULA preimplantation mammalian embryo that develops from a 32-cell stage into a fluid-filled hollow ball of over a hundred cells. A blastocyst has two distinctive tissues. The outer layer of trophoblasts gives rise to extra-embryonic tissues. The inner cell mass gives rise to the embryonic disc and eventual embryo proper.
The technique of maintaining or growing mammalian EMBRYOS in vitro. This method offers an opportunity to observe EMBRYONIC DEVELOPMENT; METABOLISM; and susceptibility to TERATOGENS.
Morphological and physiological development of EMBRYOS.
Endometrial implantation of EMBRYO, MAMMALIAN at the BLASTOCYST stage.
An early embryo that is a compact mass of about 16 BLASTOMERES. It resembles a cluster of mulberries with two types of cells, outer cells and inner cells. Morula is the stage before BLASTULA in non-mammalian animals or a BLASTOCYST in mammals.
The transfer of mammalian embryos from an in vivo or in vitro environment to a suitable host to improve pregnancy or gestational outcome in human or animal. In human fertility treatment programs, preimplantation embryos ranging from the 4-cell stage to the blastocyst stage are transferred to the uterine cavity between 3-5 days after FERTILIZATION IN VITRO.
An assisted reproductive technique that includes the direct handling and manipulation of oocytes and sperm to achieve fertilization in vitro.
The cluster of cells inside a blastocyst. These cells give rise to the embryonic disc and eventual embryo proper. They are pluripotent EMBRYONIC STEM CELLS capable of yielding many but not all cell types in a developing organism.
Morphological and physiological development of EMBRYOS or FETUSES.
The earliest developmental stage of a fertilized ovum (ZYGOTE) during which there are several mitotic divisions within the ZONA PELLUCIDA. Each cleavage or segmentation yields two BLASTOMERES of about half size of the parent cell. This cleavage stage generally covers the period up to 16-cell MORULA.
The status during which female mammals carry their developing young (EMBRYOS or FETUSES) in utero before birth, beginning from FERTILIZATION to BIRTH.
Preservation of cells, tissues, organs, or embryos by freezing. In histological preparations, cryopreservation or cryofixation is used to maintain the existing form, structure, and chemical composition of all the constituent elements of the specimens.
The entity of a developing mammal (MAMMALS), generally from the cleavage of a ZYGOTE to the end of embryonic differentiation of basic structures. For the human embryo, this represents the first two months of intrauterine development preceding the stages of the FETUS.
The fertilized OVUM resulting from the fusion of a male and a female gamete.
The transformation of a liquid to a glassy solid i.e., without the formation of crystals during the cooling process.
Delay in the attachment and implantation of BLASTOCYST to the uterine ENDOMETRIUM. The blastocyst remains unattached beyond the normal duration thus delaying embryonic development.
The formation of one or more genetically identical organisms derived by vegetative reproduction from a single cell. The source nuclear material can be embryo-derived, fetus-derived, or taken from an adult somatic cell.
Methods of maintaining or growing biological materials in controlled laboratory conditions. These include the cultures of CELLS; TISSUES; organs; or embryo in vitro. Both animal and plant tissues may be cultured by a variety of methods. Cultures may derive from normal or abnormal tissues, and consist of a single cell type or mixed cell types.
Cells lining the outside of the BLASTOCYST. After binding to the ENDOMETRIUM, trophoblasts develop into two distinct layers, an inner layer of mononuclear cytotrophoblasts and an outer layer of continuous multinuclear cytoplasm, the syncytiotrophoblasts, which form the early fetal-maternal interface (PLACENTA).
Undifferentiated cells resulting from cleavage of a fertilized egg (ZYGOTE). Inside the intact ZONA PELLUCIDA, each cleavage yields two blastomeres of about half size of the parent cell. Up to the 8-cell stage, all of the blastomeres are totipotent. The 16-cell MORULA contains outer cells and inner cells.
Female germ cells derived from OOGONIA and termed OOCYTES when they enter MEIOSIS. The primary oocytes begin meiosis but are arrested at the diplotene state until OVULATION at PUBERTY to give rise to haploid secondary oocytes or ova (OVUM).
Methods of implanting a CELL NUCLEUS from a donor cell into an enucleated acceptor cell.
Embryonic and fetal development that takes place in an artificial environment in vitro.
A unisexual reproduction without the fusion of a male and a female gamete (FERTILIZATION). In parthenogenesis, an individual is formed from an unfertilized OVUM that did not complete MEIOSIS. Parthenogenesis occurs in nature and can be artificially induced.
A tough transparent membrane surrounding the OVUM. It is penetrated by the sperm during FERTILIZATION.
The potential of the FETUS to survive outside the UTERUS after birth, natural or induced. Fetal viability depends largely on the FETAL ORGAN MATURITY, and environmental conditions.
The hollow thick-walled muscular organ in the female PELVIS. It consists of the fundus (the body) which is the site of EMBRYO IMPLANTATION and FETAL DEVELOPMENT. Beyond the isthmus at the perineal end of fundus, is CERVIX UTERI (the neck) opening into VAGINA. Beyond the isthmi at the upper abdominal end of fundus, are the FALLOPIAN TUBES.
Any liquid or solid preparation made specifically for the growth, storage, or transport of microorganisms or other types of cells. The variety of media that exist allow for the culturing of specific microorganisms and cell types, such as differential media, selective media, test media, and defined media. Solid media consist of liquid media that have been solidified with an agent such as AGAR or GELATIN.
Domesticated bovine animals of the genus Bos, usually kept on a farm or ranch and used for the production of meat or dairy products or for heavy labor.
An assisted fertilization technique consisting of the microinjection of a single viable sperm into an extracted ovum. It is used principally to overcome low sperm count, low sperm motility, inability of sperm to penetrate the egg, or other conditions related to male infertility (INFERTILITY, MALE).
The ratio of the number of conceptions (CONCEPTION) including LIVE BIRTH; STILLBIRTH; and fetal losses, to the mean number of females of reproductive age in a population during a set time period.
Occurrence or induction of release of more ova than are normally released at the same time in a given species. The term applies to both animals and humans.
Any of the processes by which nuclear, cytoplasmic, or intercellular factors influence the differential control of gene action during the developmental stages of an organism.
A colorless, odorless, viscous dihydroxy alcohol. It has a sweet taste, but is poisonous if ingested. Ethylene glycol is the most important glycol commercially available and is manufactured on a large scale in the United States. It is used as an antifreeze and coolant, in hydraulic fluids, and in the manufacture of low-freezing dynamites and resins.
A pair of highly specialized muscular canals extending from the UTERUS to its corresponding OVARY. They provide the means for OVUM collection, and the site for the final maturation of gametes and FERTILIZATION. The fallopian tube consists of an interstitium, an isthmus, an ampulla, an infundibulum, and fimbriae. Its wall consists of three histologic layers: serous, muscular, and an internal mucosal layer lined with both ciliated and secretory cells.
Methods used to induce premature oocytes, that are maintained in tissue culture, to progress through developmental stages including to a stage that is competent to undergo FERTILIZATION.
An individual that contains cell populations derived from different zygotes.
Substances that provide protection against the harmful effects of freezing temperatures.
The techniques used to select and/or place only one embryo from FERTILIZATION IN VITRO into the uterine cavity to establish a singleton pregnancy.
Genetically identical individuals developed from brother and sister matings which have been carried out for twenty or more generations, or by parent x offspring matings carried out with certain restrictions. All animals within an inbred strain trace back to a common ancestor in the twentieth generation.
The mucous membrane lining of the uterine cavity that is hormonally responsive during the MENSTRUAL CYCLE and PREGNANCY. The endometrium undergoes cyclic changes that characterize MENSTRUATION. After successful FERTILIZATION, it serves to sustain the developing embryo.
The event that a FETUS is born alive with heartbeats or RESPIRATION regardless of GESTATIONAL AGE. Such liveborn is called a newborn infant (INFANT, NEWBORN).
An acyclic state that resembles PREGNANCY in that there is no ovarian cycle, ESTROUS CYCLE, or MENSTRUAL CYCLE. Unlike pregnancy, there is no EMBRYO IMPLANTATION. Pseudopregnancy can be experimentally induced to form DECIDUOMA in the UTERUS.
The number of CELLS of a specific kind, usually measured per unit volume or area of sample.
An octamer transcription factor that is expressed primarily in totipotent embryonic STEM CELLS and GERM CELLS and is down-regulated during CELL DIFFERENTIATION.
The fusion of a spermatozoon (SPERMATOZOA) with an OVUM thus resulting in the formation of a ZYGOTE.
Cells propagated in vitro in special media conducive to their growth. Cultured cells are used to study developmental, morphologic, metabolic, physiologic, and genetic processes, among others.
The performance of dissections, injections, surgery, etc., by the use of micromanipulators (attachments to a microscope) that manipulate tiny instruments.
The outer of the three germ layers of an embryo.
Cells derived from the BLASTOCYST INNER CELL MASS which forms before implantation in the uterine wall. They retain the ability to divide, proliferate and provide progenitor cells that can differentiate into specialized cells.
Determination of the nature of a pathological condition or disease in the OVUM; ZYGOTE; or BLASTOCYST prior to implantation. CYTOGENETIC ANALYSIS is performed to determine the presence or absence of genetic disease.
The granulosa cells of the cumulus oophorus which surround the OVUM in the GRAAFIAN FOLLICLE. At OVULATION they are extruded with OVUM.
Any of various animals that constitute the family Suidae and comprise stout-bodied, short-legged omnivorous mammals with thick skin, usually covered with coarse bristles, a rather long mobile snout, and small tail. Included are the genera Babyrousa, Phacochoerus (wart hogs), and Sus, the latter containing the domestic pig (see SUS SCROFA).
The process of bearing developing young (EMBRYOS or FETUSES) in utero in non-human mammals, beginning from FERTILIZATION to BIRTH.
The inner of the three germ layers of an embryo.
Early pregnancy loss during the EMBRYO, MAMMALIAN stage of development. In the human, this period comprises the second through eighth week after fertilization.
An INTERLEUKIN-6 related cytokine that exhibits pleiotrophic effects on many physiological systems that involve cell proliferation, differentiation, and survival. Leukemia inhibitory factor binds to and acts through the lif receptor.
Elements of limited time intervals, contributing to particular results or situations.
Results of conception and ensuing pregnancy, including LIVE BIRTH; STILLBIRTH; SPONTANEOUS ABORTION; INDUCED ABORTION. The outcome may follow natural or artificial insemination or any of the various ASSISTED REPRODUCTIVE TECHNIQUES, such as EMBRYO TRANSFER or FERTILIZATION IN VITRO.
The process of germ cell development in the female from the primordial germ cells through OOGONIA to the mature haploid ova (OVUM).
The three primary germinal layers (ECTODERM; ENDODERM; and MESODERM) developed during GASTRULATION that provide tissues and body plan of a mature organism. They derive from two early layers, hypoblast and epiblast.
Progressive restriction of the developmental potential and increasing specialization of function that leads to the formation of specialized cells, tissues, and organs.
Liquid components of living organisms.
The major progestational steroid that is secreted primarily by the CORPUS LUTEUM and the PLACENTA. Progesterone acts on the UTERUS, the MAMMARY GLANDS and the BRAIN. It is required in EMBRYO IMPLANTATION; PREGNANCY maintenance, and the development of mammary tissue for MILK production. Progesterone, converted from PREGNENOLONE, also serves as an intermediate in the biosynthesis of GONADAL STEROID HORMONES and adrenal CORTICOSTEROIDS.
Mature male germ cells derived from SPERMATIDS. As spermatids move toward the lumen of the SEMINIFEROUS TUBULES, they undergo extensive structural changes including the loss of cytoplasm, condensation of CHROMATIN into the SPERM HEAD, formation of the ACROSOME cap, the SPERM MIDPIECE and the SPERM TAIL that provides motility.
A polymer prepared from polyvinyl acetates by replacement of the acetate groups with hydroxyl groups. It is used as a pharmaceutic aid and ophthalmic lubricant as well as in the manufacture of surface coatings artificial sponges, cosmetics, and other products.
Methods for maintaining or growing CELLS in vitro.
The division of a ZYGOTE into two parts, each of which is capable of further development.
Liquids transforming into solids by the removal of heat.
RNA sequences that serve as templates for protein synthesis. Bacterial mRNAs are generally primary transcripts in that they do not require post-transcriptional processing. Eukaryotic mRNA is synthesized in the nucleus and must be exported to the cytoplasm for translation. Most eukaryotic mRNAs have a sequence of polyadenylic acid at the 3' end, referred to as the poly(A) tail. The function of this tail is not known for certain, but it may play a role in the export of mature mRNA from the nucleus as well as in helping stabilize some mRNA molecules by retarding their degradation in the cytoplasm.
The hormone-responsive glandular layer of ENDOMETRIUM that sloughs off at each menstrual flow (decidua menstrualis) or at the termination of pregnancy. During pregnancy, the thickest part of the decidua forms the maternal portion of the PLACENTA, thus named decidua placentalis. The thin portion of the decidua covering the rest of the embryo is the decidua capsularis.
The developmental history of specific differentiated cell types as traced back to the original STEM CELLS in the embryo.
Procedures to obtain viable OOCYTES from the host. Oocytes most often are collected by needle aspiration from OVARIAN FOLLICLES before OVULATION.
The process by which a tissue or aggregate of cells is kept alive outside of the organism from which it was derived (i.e., kept from decay by means of a chemical agent, cooling, or a fluid substitute that mimics the natural state within the organism).
Cells that can give rise to cells of the three different GERM LAYERS.
Within a eukaryotic cell, a membrane-limited body which contains chromosomes and one or more nucleoli (CELL NUCLEOLUS). The nuclear membrane consists of a double unit-type membrane which is perforated by a number of pores; the outermost membrane is continuous with the ENDOPLASMIC RETICULUM. A cell may contain more than one nucleus. (From Singleton & Sainsbury, Dictionary of Microbiology and Molecular Biology, 2d ed)
A species of SWINE, in the family Suidae, comprising a number of subspecies including the domestic pig Sus scrofa domestica.
A proteolytic enzyme obtained from Streptomyces griseus.
Clinical and laboratory techniques used to enhance fertility in humans and animals.
The injection of very small amounts of fluid, often with the aid of a microscope and microsyringes.

An ultrastructural study of implantation in the golden hamster. II. Trophoblastic invasion and removal of the uterine epithelium. (1/4036)

Sixty six implantation sites from 18 golden hamsters were examined with light and electron microscopy between 4 and 5 1/2 days of pregnancy (post-ovulation). At 4 days some blastocysts began to invade the uterine epithelium, with trophoblastic processes penetrating and engulfing portions of the uterine epithelium. The majority of epithelial cells appeared normal before invasion, although at two implantation sites three or four adjoining epithelial cells were necrotic before penetration by the trophoblast. In general the epithelial cells were degenerating at the time the trophoblast invaded the epithelium. Inclusions, representing portions of the engulfed epithelium, and varying in size and electron density, were present throughout the invading trophoblast cells at 4 1/2 and 5 days of pregnancy. At 5 1/2 days the uterine epithelium had disappeared and the embryo was now almost completely surrounded by blood lacunae.  (+info)

Ontogeny of expression of a receptor for platelet-activating factor in mouse preimplantation embryos and the effects of fertilization and culture in vitro on its expression. (2/4036)

Platelet-activating factor (PAF; 1-o-alkyl-2-acetyl-sn-glycero-3-phosphocholine) is a potent ether phospholipid. It is one of the preimplantation embryo's autocrine growth/survival factors. It may act via a G protein-linked receptor on the embryo; however, the evidence for this is conflicting. The recent description of the intracellular form of the PAF:acetlyhydrolase enzyme as having structural homology with G proteins and Ras also suggests this as a potential intracellular receptor/transducer for PAF. This study used reverse transcription-polymerase chain reaction to examine the ontogeny of expression of the genes for these proteins in the oocyte and preimplantation-stage embryo. Transcripts for the G protein-linked PAF receptor were detected in the late 2-cell-stage embryo and in all stages from the 4-cell stage to blastocysts. They were also present in unfertilized oocytes and newly fertilized zygotes but only at relatively low levels. The incidence of expression was generally low and variable in late zygotes and early 2-cell embryos. Expression past the 2-cell stage was alpha-amanitin sensitive. The results indicated that mRNA for this receptor is a maternal transcript that was degraded during the zygote-2-cell stage. New expression of the receptor transcript required activation of the zygotic genome. Fertilization of embryos in vitro caused this transcript not to be expressed in the zygote. Culture of zygotes (irrespective of their method of fertilization) caused expression from the zygotic genome to be retarded by more than 24 h. This retardation did not occur if culture commenced at the 2-cell stage. The transcripts for the subunits of intracellular PAF:acetylhydrolase were not detected in oocytes or at any stage of embryo development examined, despite their being readily detected in control tissue. This study confirms the presence of the G protein-linked PAF receptor in the 2-cell embryo and describes for the first time its normal pattern of expression during early development. The adverse effects of in vitro fertilization (IVF) and embryo culture on the expression of this transcript may be a contributing factor for the poor viability of embryos produced in this manner. The reduced expression of PAF-receptor mRNA following IVF predicts that such embryos may have a deficiency in autocrine stimulation and also suggests that supplementation of growth media with exogenous PAF would be only partially beneficial. The effect of IVF and culture may also explain the conflicting literature.  (+info)

X inactive-specific transcript (Xist) expression and X chromosome inactivation in the preattachment bovine embryo. (3/4036)

Expression of the X inactive-specific transcript (Xist) is thought to be essential for the initiation of X chromosome inactivation and dosage compensation during female embryo development. In the present study, we analyzed the patterns of Xist transcription and the onset of X chromosome inactivation in bovine preattachment embryos. Reverse transcription-polymerase chain reaction (RT-PCR) revealed the presence of Xist transcripts in all adult female somatic tissues evaluated. In contrast, among the male tissues examined, Xist expression was detected only in testis. No evidence for Xist transcription was observed after a single round of RT-PCR from pools of in vitro-derived embryos at the 2- to 4-cell stage. Xist transcripts were detected as a faint amplicon at the 8-cell stage initially, and consistently thereafter in all stages examined up to and including the expanded blastocyst stage. Xist transcripts, however, were subsequently detected from the 2-cell stage onward after nested RT-PCR. Preferential [3H]thymidine labeling indicative of late replication of one of the X chromosomes was noted in female embryos of different developmental ages as follows: 2 of 7 (28.5%) early blastocysts, 6 of 13 (46.1%) blastocysts, 8 of 11 (72.1%) expanded blastocysts, and 14 of 17 (77.7%) hatched blastocysts. These results suggest that Xist expression precedes the onset of late replication in the bovine embryo, in a pattern compatible with a possible role of bovine Xist in the initiation of X chromosome inactivation.  (+info)

Induction of Ig light chain gene rearrangement in heavy chain-deficient B cells by activated Ras. (4/4036)

During B cell development, rearrangement and expression of Ig heavy chain (HC) genes promote development and expansion of pre-B cells accompanied by the onset of Ig light chain (LC) variable region gene assembly. To elucidate the signaling pathways that control these events, we have tested the ability of activated Ras expression to promote B cell differentiation to the stage of LC gene rearrangement in the absence of Ig HC gene expression. For this purpose, we introduced an activated Ras expression construct into JH-deleted embryonic stem cells that lack the ability to assemble HC variable region genes and assayed differentiation potential by recombination activating gene (RAG) 2-deficient blastocyst complementation. We found that activated Ras expression induces the progression of B lineage cells beyond the developmental checkpoint ordinarily controlled by mu HC. Such Ras/JH-deleted B cells accumulate in the periphery but continue to express markers associated with precursor B cells including RAG gene products. These peripheral Ras/JH-deleted B cell populations show extensive Ig LC gene rearrangement but maintain an extent of kappa LC gene rearrangement and a preference for kappa over lambda LC gene rearrangement similar to that of wild-type B cells. We discuss these findings in the context of potential mechanisms that may regulate Ig LC gene rearrangement.  (+info)

In-vitro fertilization and culture of mouse embryos in vitro significantly retards the onset of insulin-like growth factor-II expression from the zygotic genome. (5/4036)

In this study, the effect of in-vitro fertilization (IVF) and culture of mouse embryos in vitro on the normal expression of insulin-like growth factor-II (IFG-II) ligand and receptor was examined. The expression of IGF-II increased in a linear fashion at least up to the 8-cell stage of development. IGF-II expression in embryos collected fresh from the reproductive tract was significantly (P < 0.001) greater than in embryos fertilized in the reproductive tract and cultured in vitro (in-situ fertilized: ISF), and its expression was further reduced (P < 0.001) in IVF embryos at all development stages tested. The expression of IGF-II was significantly (P < 0.001) lower when embryos were cultured individually in 100 microl drops compared with culture in groups of 10 in 10 microl drops of medium. The addition of platelet activating factor to culture medium partially overcame this density-dependent decline of expression. Culture of ISF and IVF zygotes also caused the onset of new IGF-II mRNA transcription from the zygotic genome to be significantly (P < 0.001) retarded, until at least the 8-cell stage of development. This effect was greater (P < 0.05) for IVF than for ISF embryos. Neither IVF nor culture had any obvious effect on IFG-II/mannose-6-phosphate receptor (IGF-IIr) mRNA expression.  (+info)

Detection of benzo[a]pyrene diol epoxide-DNA adducts in embryos from smoking couples: evidence for transmission by spermatozoa. (6/4036)

Tobacco smoking is deleterious to reproduction. Benzo[a]pyrene (B[a]P) is a potent carcinogen in cigarette smoke. Its reactive metabolite induces DNA-adducts, which can cause mutations. We investigated whether B[a]P diol epoxide (BPDE) DNA adducts are detectable in preimplantation embryos in relation to parental smoking. A total of 17 couples were classified by their smoking habits: (i) both partners smoke; (ii) wife non-smoker, husband smokes; and (iii) both partners were non-smokers. Their 27 embryos were exposed to an anti-BPDE monoclonal antibody that recognizes BPDE-DNA adducts. Immunostaining was assessed in each embryo and an intensity score was calculated for embryos in each smoking group. The proportion of blastomeres which stained was higher for embryos of smokers than for non-smokers (0.723 versus 0.310). The mean intensity score was also higher for embryos of smokers (1.40+/-0.28) than for non-smokers (0.38+/-0.14; P = 0.015), but was similar for both types of smoking couples. The mean intensity score was positively correlated with the number of cigarettes smoked by fathers (P = 0.02). Increased mean immunostaining in embryos from smokers, relative to non-smokers, indicates a relationship with parental smoking. The similar levels of immunostaining in embryos from both types of smoking couples suggest that transmission of modified DNA is mainly through spermatozoa. We confirmed paternal transmission of modified DNA by detection of DNA adducts in spermatozoa of a smoker father and his embryo.  (+info)

Co-expression of cytokeratins and vimentin by highly invasive trophoblast in the white-winged vampire bat, Diaemus youngi, and the black mastiff bat, Molossus ater, with observations on intermediate filament proteins in the decidua and intraplacental trophoblast. (7/4036)

Histological and immunocytochemical studies of gravid reproductive tracts obtained from the white-winged vampire bat (Diaemus youngi) and the black mastiff bat (Molossus ater) have established that both species develop unusually invasive trophoblast. This is released by the developing discoidal haemochorial placenta, expresses both cytokeratins and vimentin, and invades the myometrium and adjacent tissues (including the ovaries) via interstitial migration within the walls of maternal blood vessels. Hence, this trophoblast is noteworthy for the extent to which it undergoes an epithelial-mesenchymal transformation. In Molossus, it originates from the cytotrophoblastic shell running along the base of the placenta, is mononuclear, and preferentially invades maternal arterial vessels serving the discoidal placenta. This trophoblast may have a role in dilatation of these vessels when the discoidal placenta becomes functional. In Diaemus, the highly invasive trophoblast appears to originate instead from a layer of syncytiotrophoblast on the periphery of the placenta is multinucleated, and vigorously invades both arterial and venous vessels. During late pregnancy, it becomes extensively branched and sends attenuated processes around many of the myometrial smooth muscle fibres. In view of its distribution, this trophoblast could have important influences upon myometrial contractility and the function of blood vessels serving the gravid tract. Other aspects of intermediate filament expression in the uteri and placentae of these bats are also noteworthy. Many of the decidual giant cells in Molossus co-express cytokeratins and vimentin, while the syncytiotrophoblast lining the placental labyrinth in Diaemus late in pregnancy expresses little cytokeratin.  (+info)

Trophectoderm differentiation in the bovine embryo: characterization of a polarized epithelium. (8/4036)

Blastocytst formation is dependent on the differentiation of a transporting epithelium, the trophectoderm, which is coordinated by the embryonic expression and cell adhesive properties of E-cadherin. The trophectoderm shares differentiative characteristics with all epithelial tissues, including E-cadherin-mediated cell adhesion, tight junction formation, and polarized distribution of intramembrane proteins, including the Na-K ATPase. The present study was conducted to characterize the mRNA expression and distribution of polypeptides encoding E-cadherin, beta-catenin, and the tight junction associated protein, zonula occludens protein 1, in pre-attachment bovine embryos, in vitro. Immunocytochemistry and gene specific reverse transcription--polymerase chain reaction methods were used. Transcripts for E-cadherin and beta-catenin were detected in embryos of all stages throughout pre-attachment development. Immunocytochemistry revealed E-cadherin and beta-catenin polypeptides evenly distributed around the cell margins of one-cell zygotes and cleavage stage embryos. In the morula, detection of these proteins diminished in the free apical surface of outer blastomeres. E-cadherin and beta-catenin became restricted to the basolateral membranes of trophectoderm cells of the blastocyst, while maintaining apolar distributions in the inner cell mass. Zonula occludens protein 1 immunoreactivity was undetectable until the morula stage and first appeared as punctate points between the outer cells. In the blastocyst, zonula occludens protein 1 was localized as a continuous ring at the apical points of trophectoderm cell contact and was undetectable in the inner cell mass. These results illustrate that the gene products encoding E-cadherin, beta-catenin and zonula occludens protein 1 are expressed and maintain cellular distribution patterns consistent with their predicted roles in mediating trophectoderm differentiation in in vitro produced bovine embryos.  (+info)

STUDY QUESTION: Is the spindle assembly checkpoint (SAC) active during human preimplantation development?. SUMMARY ANSWER: Mitotic spindle disruption during mitosis activates the SAC from at least Day 3 of human preimplantation development, but this does not lead to apoptosis until Day 5.. WHAT IS KNOWN ALREADY: Human preimplantation embryos frequently acquire chromosomal abnormalities, but the mechanisms behind this are poorly understood. It has been speculated that a dysfunctional SAC could be responsible. Although research has shown that the SAC components are present during early human development, functional studies are lacking.. STUDY DESIGN, SIZE, DURATION: In vitro study using human preimplantation embryos in a university research laboratory. We studied a total of 38 Day-3, 38 Day-4, 29 Day-5 and 21 Day-6 human preimplantation embryos, donated for research, during 16 h of incubation.. PARTICIPANT/MATERIALS, SETTING, METHODS: We cultured human preimplantation embryos overnight in a ...
TY - JOUR. T1 - Brain and sperm cell surface antigen (NS-4) on preimplantation mouse embryos. AU - Solter, Davor. AU - Camartin, Melitta. PY - 1976/1/1. Y1 - 1976/1/1. N2 - Antiserum prepared in rabbit against 4-day-old mouse cerebellum (anti-NS-4 serum) reacts in the complement-mediated cytotoxicity test with unfertilized and fertilized mouse eggs, cleavage stage embryos, and cells of the trophoblast and inner cell mass of the mouse blastocyst. This activity is specifically removed by absorption of antiserum with adult mouse brain and epididymal sperm but not with adult liver, spleen, kidney, and thymocytes. The antiserum reacts most strongly with cells of the trophoblast and inner cell mass and, in order of decreasing reactivity, with four- to eight-cell stage embryos, zygotes, unfertilized eggs, and two-cell stage embryos.. AB - Antiserum prepared in rabbit against 4-day-old mouse cerebellum (anti-NS-4 serum) reacts in the complement-mediated cytotoxicity test with unfertilized and fertilized ...
Introduction: Cryopreservation is possible for all stages of pre-implantation embryos. It has been reported that survival rate of blastocyst is comparably lower than other stages. There is a high volume of fluid in blastocoel cavity that can be a good ground for ice crystals formation, resulting in damage to the cell structure. In this study, the effects of artificial collapseand reduction of the fluid volume in blastocyst cavity before vitrification process on the survival rate and quality of blastocysts were assessed by estimating the expression levels of Oct4, Nanog, Sox2, and Klf4.. Materials & Methods: Mouse blastocysts divided into 5 groups including: A) Vitrified- thawed blastocysts, B) Vitrified- thawed blastocysts after artificial collapse, C) Collapsed blastocysts, D) Immersed blastocysts in vitrification/warming solutions and E) Fresh blastocysts as control group. The survival and hatching rate of embryos were evaluated and the expression of pluripotency-specific genes was assessed by ...
The degradation of maternal proteins is one of the most important events during early development, and it is presumed to be essential for embryonic genome activation (EGA), but the precise mechanism is still not known. It is thought that a large proportion of the degradation of maternal proteins is mediated by the ubiquitin-proteolytic system. In this study we focused on the expression of the Skp1-Cullin1-F-box (SCF) complex, a modular RING-type E3 ubiquitin-ligase, during bovine preimplantation development. The complex consists of three invariable components--Cul1, Skp1, Rbx1 and F-box protein, which determines the substrate specificity. The protein level and mRNA expression of all three invariable members were determined. Cul1 and Skp1 mRNA synthesis was activated at early embryonic stages, at the 4c and early 8c stage, respectively, which suggests that these transcripts are necessary for preparing the embryo for EGA. CUL1 protein level increased from MII to the morula stage, with a ...
Calcitonin secretion in the pregnant uterus is tightly regulated by the ovarian hormones, estrogen and progesterone, which limit its expression to a brief period preceding blastocyst implantation. The binding of calcitonin to a G protein-coupled receptor activates adenylate cyclase and elevates cytosolic Ca2+ levels. The acceleration of preimplantation embryonic development that is known to occur upon elevation of intracellular Ca2+ prompted an investigation into calcitonin regulation of blastocyst differentiation. Using reverse transcription and the polymerase chain reaction to estimate the relative abundance of calcitonin receptor mRNA, a 25-fold accumulation of the splice variant, CR-1a, was observed in embryos between the 1-cell and 8-cell stages. Cytosolic free Ca2+ levels were rapidly elevated in embryos at the 4-cell to blastocyst stages after exposure to 10 nM calcitonin. Blastocysts treated for 30 minutes with 10 nM calcitonin differentiated in vitro at an accelerated rate, as assessed ...
Totipotent non-committed inner cell mass (ICM) cells from human blastocysts, if demonstrated to be capable of proliferating in vitro without differentiation, will have several beneficial uses, not only in the treatment of neurodegenerative and genetic disorders, but also as a model in studying the events involved in embryogenesis and genomic manipulation. Nine patients admitted to an in-vitro fertilization programme donated 21 spare embryos for this study. All 21 embryos were grown from the 2-pronuclear until blastocyst stages on a human tubal epithelial monolayer in commercial Earles medium (Medicult, Denmark) supplemented with 10% human serum. The medium was changed after blastocyst formation to Changs medium supplemented with 1000 units/ml of human leukaemia inhibitory factor (HLIF) and the embryos left undisturbed for 72 h to allow the hatched ICM and trophoblast to attach to the feeder monolayer. Nineteen of the 21 embryos from nine patients produced healthy ICM lumps which could be ...
Selective labelling of polar trophectoderm cells in early mouse blastocysts has allowed the fate of polar cells to be followed during in vitro and in vivo blastocyst development. Results show that there is cell movement from polar to mural regions as blastocysts grow. This indicates that trophectode …
Immunosurgery is a useful technique for the isolation of inner cell masses from murine blastocysts. Conventionally, rabbit antisera made ad hoc against murine splenic or fetal cells or fibroblasts have been used as antibody sources. We investigated the feasibility of using commercially available rabbit antiserum to murine erythrocytes (anti-RBC) and compared it with rabbit antiserum generated ad hoc to murine L-cells (anti-L-cell). Our results indicate that anti-RBC is at least as effective as anti-L-cell serum for the immunosurgical isolation of inner cell masses, which became either miniblastocysts (later forming outgrowths) or embryoid bodies (undergoing ectoderm-endodermlike differentiation within 48 h). Because anti-RBC is commercially available, the technical modification described herein increases the accessibility of the immunosurgical protocol for the isolation of murine inner cell masses.
The accumulation of substrate carbon by mouse embryos was measured following incubation in U-14C-glucose. Following a 30-min incubation period 273 × 10-14 and 301×10-14g atoms of substrate carbon per embryo were found in 2- and 8-cell embryos respectively. By comparison, the figures for unfertilized and fertilized ova were 14 × 10 -14 and 45 × 10-14 g atoms of substrate carbon.. The intracellular concentration of substrate carbon was timedependent in both 2- and 8-cell embryos. After an 80-min incubation, substrate carbon in the 8-cell embryo was almost double that in the 2-cell embryo. Accumulation did not occur during incubations at 5° C and there was competition between glucose and galactose for uptake. The results are discussed in relation to the energy requirements of the developing zygote. ...
Fibronectin 1 (FN1), a glycoprotein component of the extracellular matrix, exerts different functions during reproductive processes such as fertilisation, gastrulation and implantation. FN1 expression has been described to increase significantly from the morula towards the early blastocyst stage, suggesting that FN1 may also be involved in early blastocyst formation. By alternative splicing at 3 defined regions, different FN1 isoforms are generated, each with a unique biological function. The analysis of the alternative FN1 splicing on the one hand and the search for candidate FN1 receptors on the other hand during early bovine embryo development may reveal more about its function during bovine preimplantation embryo development. RT-qPCR quantification of the FN1 splice isoforms in oocytes, embryos, cumulus cells and adult tissue samples revealed a large variation in overall FN1 expression and in splice variant expression. Moreover, two new FN1 transcript variants were identified, the first one
Generation Functional Lungs Conditional Blastocyst Complementation Using Pluripotent PubMed Journal Articles published on BioPortfolio | BioPortfolio
Early development and implantation of the embryo.. A. The zygote stage begins upon fertilization of the secondary oocyte by the sperm. The zygote contains both pro-nuclei and is contained within the zona pellucida, until the blastocyst stage. B. The morula stage. Following compaction and symmetrical cleavage divisions of the blastomeres (the cells of the early embryo), the embryo contains 8 (early morula) to 32 cells (morula). The inner cells will give rise to the inner cell mass, whereas the outer cells will give rise to the trophoblast, which forms a cavity called the blastocoele cavity. C. The blastocyst stage. The developing embryo is defined as a blastocyst from the appearance of the blastocoele cavity, and now contains two cell populations- the surrounding outer trophoblast cells, and the inner cell mass cells, located at one side of the inner cavity. The portion of the trophoblast nearest to the inner cell mass is called the polar trophoblast (embryonic pole) and the portion of the ...
Embryos cryopreserved after reaching blastocyst stage on day 7 were compared to those cryopreserved on days 5 and 6. Day 7 blastocysts have lower but clinically important pregnancy rates.
Oct4 is required for lineage priming in the developing inner cell mass of the mouse blastocyst[4] The transcription factor Oct4 is required in vitro for establishment and maintenance of embryonic stem cells and for reprogramming somatic cells to pluripotency. In vivo, it prevents the ectopic differentiation of early embryos into trophoblast. Here, we further explore the role of Oct4 in blastocyst formation and specification of epiblast versus primitive endoderm lineages using conditional genetic deletion. Experiments involving mouse embryos deficient for both maternal and zygotic Oct4 suggest that it is dispensable for zygote formation, early cleavage and activation of Nanog expression. Nanog protein is significantly elevated in the presumptive inner cell mass of Oct4 null embryos, suggesting an unexpected role for Oct4 in attenuating the level of Nanog, which might be significant for priming differentiation during epiblast maturation. Induced deletion of Oct4 during the morula to blastocyst ...
Hewitson, L.C., Leese, H.J. (1993) Energy metabolism of the trophectoderm and inner cell mass of the mouse blastocyst. J Exp Zool. 267:337-343. Hewitson, L.C., Martin, K.L., Leese, H.J. (1996) Effects of metabolic inhibitors on mouse preimplantation embryo development and the energy metabolism of isolated inner cell masses. Mol Reprod Dev. 43:323-330. Hewitson, L., Simerly, C., Tengowski, M.W, Sutovsky, P., Navara, C,S., Haavisto, A.J. and Schatten, G. (1996) Microtubule and chromatin configurations during rhesus intracytoplasmic sperm injection: Successes and Failures. Biol. Reprod. 55:271-280. Hewitson, L., Haavisto A, Simerly C, Jones J and Schatten G (1997) Microtubule organization and chromatin configurations in hamster oocytes during fertilization, parthenogenetic activation and after insemination with human sperm. Biol.Reprod. 57: 967-975. Hewitson L, Takahashi D, Dominko T, Simerly C, and Schatten G. (1998) Fertilization and embryo development to blastocysts after intracytoplasmic sperm ...
A sequential study was made of non-delayed and of experimentally delayed rat blastocysts. Alterations in shape, axis lengths and area of non-delayed blastocysts first appeared on the afternoon of Day 5 of pregnancy. Similar changes were observed in delayed blastocysts beginning 12 hr after the simultaneous administration of oestrone and progesterone. In both cases implantation occurred within the subsequent 24 hr. In contrast, delayed blastocysts maintained only with progesterone were markedly larger than normal but failed to demonstrate typical pre-implantation changes or to implant. Therefore it was concluded that changes necessary for and indicative of impending implantation are induced by the synergistic action of oestrone and progesterone on the blastocyst.. Blastocysts from non-delayed and delayed, oestrone-treated animals were consistently free of their zonae pellucidae at least 18 hr prior to implantation-6 hr after pre-implantation changes were initially noted. Progesterone, although it ...
Preimplantation mouse embryos of many stresses become arrested at the 2-cell stage if the osmolarity of culture medium that normally supports development to blastocysts is raised to approximately that of their normal physiological environment in the oviduct. cultured at 310 mOsM progressed through the S phase, and zygotic genome activation markers were expressed. However, most embryos failed to initiate the M phase, as evidenced by intact nuclei with decondensed chromosomes, low M-phase promoting factor activity, and an inactive form of CDK1, although a few blastomeres were arrested in metaphase. Thus, embryos become arrested late in the G2 stage of the second embryonic cell cycle when stressed by physiological osmolarity in the absence of organic osmolytes. was detected using immunocytochemistry as explained previously [22] with minor modifications. All procedures were performed at 37C unless normally noted. Briefly, embryos were incubated in 400 nM MitoTracker (MitoTracker Red CMXRos; ...
Measuring gene expression in individual cells is crucial for understanding the gene regulatory network controlling human embryonic development. Here we apply single-cell RNA sequencing (RNA-Seq) analysis to 124 individual cells from human preimplantation embryos and human embryonic stem cells (hESCs …
The cell lineage of the mouse was studied from the 2-cell stage to the blastocyst. Lineage to the 8-cell stage was followed under the microscope. Each cell from the 2-cell stage divided to form two daughter cells which remained attached. Subsequently, these two daughters each produced two descendants; one of these descendants regularly lay deep in the structure of the embryo while the other was peripheral. Lineage to the blastocyst was followed by injecting oil drops into cells at the 8-cell stage, and then following the segregation of these drops into the inner cell mass and trophectoderm. Between the 8-cell stage and the blastocyst, the deep cells contributed more frequently to the inner cell mass than did the peripheral cells.. ...
Treatment of in vitro matured bovine oocytes with colcemid results in a membrane protrusion that contains maternal chromosomes, which can be easily removed by aspiration. Four experiments were designed to evaluate the overall and temporal effects of conditioned medium (CM) by bovine cumulus cells on development of nuclear transfer (NT) bovine embryos and to examine the chromosomal composition and allocation of inner cell mass (ICM) and trophectoderm (TE) of the subsequent blastocysts. The nuclear transfer embryos were cultured in various CR1aa media conditioned by preculture with bovine cumulus cells. Development to the blastocyst stage in BSA-containing CM (BCM) and serum-containing CM (SCM) were similar to co-culture group (24-30%). The 24 hr-conditioned BCM yielded higher blastocyst development than 48 and 72 hr-conditioned BCM. Temporary exposure of embryos to BCM and SCM followed by CR1aa was also studied. Morula and blastocyst development were not different among the groups cultured in BCM for 72,
Bangalore Infertility Treatments center,The couple undergoing for IVF/ICSI have more embryos.Blastocyst Culture,Technique in IVF to select the best embryo.
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In humans, reproduction is considered a relatively inefficient process, when compared with other mammalian species and the chance of achieving a spontaneous pregnancy after timed intercourse is at the most 20-30%. Chromosome segregation errors are a well-known inherent feature of cell division in human preimplantation embryos produced by in vitro ... read more fertilization (IVF). The occurrence of such errors, which results in embryos containing chromosomally abnormal (aneuploid) cells, is believed to be the main cause for the reported inefficiency of human reproduction, as it may lead to pre-clinical pregnancy losses. In this thesis we start by evaluating the impact of ovarian stimulation administrated to patients undergoing IVF on the development of IVF-derived human embryos. We conclude that although the use of ovarian stimulation is considered relatively safe, further studies are needed to increase the knowledge on this subject and reduce potential effects on embryo development and ...
Computational analysis of single-cell transcriptomics data elucidates the stabilization of Oct4 expression in the E3.25 mouse preimplantation embryo [6] Our computational analysis focuses on the 32- to 64-cell mouse embryo transition, Embryonic day (3.25), whose study in literature is concentrated mainly on the search for an early onset of the second cell-fate decision, the specification of the inner cell mass (ICM) to primitive endoderm (PE) and epiblast (EPI). We analysed single-cell (sc) microarray transcriptomics data from E3.25 using Hierarchical Optimal k-Means (HOkM) clustering, and identified two groups of ICM cells: a group of cells from embryos with less than 34 cells (E3.25-LNCs), and another group of cells from embryos with more than 33 cells (E3.25-HNCs), corresponding to two developmental stages. Although we found massive underlying heterogeneity in the ICM cells at E3.25-HNC with over 3,800 genes with transcriptomics bifurcation, many of which are PE and EPI markers, we showed ...
The expression of microRNAs (miRNAs) is essential for the proper development of the mammalian embryo. A maternal exposure to endocrine disrupting chemicals during preimplantation bears the potential for transgenerational inheritance of disease through the epigenetic perturbation of the developing embryo. A comprehensive assembly of embryo-specific miRNAs and respective isoforms (isomiR) is lacking to date. We aimed at revealing the sex-specific miRNA expression profile of single porcine blastocysts developing in gilts orally exposed to exogenous estradiol-17 β (E2). Therefore we analyzed the miRNA profile specifically focusing on isomiRs and potentially embryo-specific miRNAs. Deep sequencing of small RNA (small RNA-seq) result in the detection of miRNA sequences mapping to known and predicted porcine miRNAs as well as novel miRNAs highly conserved in human and cattle. A set of highly abundant miRNAs and a large number of rarely expressed miRNAs were identified by using a small RNA analysis pipeline,
The molecular regulation of mammalian peri-implantation development is complex and difficult to study in vivo. We successfully cultured hamster blastocysts through hatching and peri-attachment stages, using a chemically defined medium, HECM-2h. Using this system, we showed that a species-specific, embryonic cysteine-like protease is involved in blastocyst hatching and that the process is modulated by growth factors. In particular, heparin binding-epidermal growth factor (HB-EGF) or leukemia inhibitory factor (LIF) enhance blastocyst hatching, and the former also improves attachment and trophoblast outgrowth. We observed interesting changing patterns of expression of mRNA and/or immunoreactive protein for EGF, HB-EGF, LIF and transforming growth factor-beta (TGF-beta) in the embryo and/or endometrial tissue, during peri-implantation development. Together, it appears that hamster blastocyst hatching, attachment and trophoblast outgrowth are regulated by autocrine and/or paracrine growth factors, ...
Blastocyst Transfer and treatment at Santati Fertility Center in Mumbai and Thane, the Indias largest independent infertility Blastocyst Transfer treatment provider
We examined the in vitro developmental competence of parthenogenetic activation (PA) oocytes activated by an electric pulse (EP) and treated with various concentrations of AZD5438 for 4 h. Treatment with 10 µM AZD5438 for 4 h significantly improved the blastocyst formation rate of PA oocytes in comparison with 0, 20, or 50 µM AZD5438 treatment (46.4% vs. 34.5%, 32.3%, and 24.0%, respectively; P , 0.05). The blastocyst formation rate was higher in the group treated with AZD5438 for 4 h than in the groups treated with AZD5438 for 2 or 6 h (42.8% vs. 38.6% and 37.2%, respectively; P , 0.05). Furthermore, 66.67% of blastocysts derived from these AZD5438-treated PA oocytes had a diploid karyotype. The blastocyst formation rate of PA and somatic cell nuclear transfer (SCNT) embryos was similar between oocytes activated by an EP and treated with 2 mM 6-dimethylaminopurine for 4 h and those activated by an EP and treated with 10 µM AZD5438 for 4 h (11.11% vs. 13.40%, P , 0.05). In addition, the level ...
We examined the in vitro developmental competence of parthenogenetic activation (PA) oocytes activated by an electric pulse (EP) and treated with various concentrations of AZD5438 for 4 h. Treatment with 10 µM AZD5438 for 4 h significantly improved the blastocyst formation rate of PA oocytes in comparison with 0, 20, or 50 µM AZD5438 treatment (46.4% vs. 34.5%, 32.3%, and 24.0%, respectively; P < 0.05). The blastocyst formation rate was higher in the group treated with AZD5438 for 4 h than in the groups treated with AZD5438 for 2 or 6 h (42.8% vs. 38.6% and 37.2%, respectively; P > 0.05). Furthermore, 66.67% of blastocysts derived from these AZD5438-treated PA oocytes had a diploid karyotype. The blastocyst formation rate of PA and somatic cell nuclear transfer (SCNT) embryos was similar between oocytes activated by an EP and treated with 2 mM 6-dimethylaminopurine for 4 h and those activated by an EP and treated with 10 µM AZD5438 for 4 h (11.11% vs. 13.40%, P > 0.05). In addition, the level ...
The aim in developing this approach to the study of implantation has been to establish a method which will permit the investigation of the process extracorporeally, thus providing conditions that are more readily controllable than those obtainable in an experimental animal. Shaffers method of organ culture has been adapted to the needs of the present investigation and it has been found that somewhat more than half of the rabbit blastocysts, explanted on strips of endometrium and incubated on them as combined explants, became implanted within 48 to 72 h. ...
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I had 2, 5-day blastocyst transferred exactly 2 weeks ago. I had a faint positive home test at 9 days post transfer. 3 days later I had some brown bloody
The preference of fertilized (IVF) and somatic cell nuclear transfer (SCNT) presumptive zygotes for different media when cultured in vitro to the blastocyst stage was evaluated in this study. The experiment comprised two zygote production methods (IVF and SCNT) × two culture media (mSOF and G1.5/G2.5) factorial design in which culture droplets that contained approximate 30 presumptive zygotes formed the experimental plots for the assessment of cleavage and blastocyst development. There were 15 to 20 replicates (culture droplets) per treatment combination. Sub-samples 30 to 41 of the blastocysts produced were assessed for cell number and cell apoptosis. A further 10 blastocysts per treatment combination were used for quantitative real-time polymerase chain reaction (RT-PCR) to evaluate the relative abundance of Hsp70 and Bax mRNA. Presumptive zygotes produced by IVF were developmentally more competent than SCNT zygotes in terms of cleavage rate (66.9 vs. 57.0%; P , 0.05) and blastocyst ...
Nilsson, B. O., Jin, M., Larsson, A. and Sundström, P. (1996), Human Autoantibodies Recognizing Human and Mouse Preimplantation Stages. American Journal of Reproductive Immunology, 36: 135-140. doi: 10.1111/j.1600-0897.1996.tb00154.x ...
The preimplantation period of development represents the highest interval of embryonic loss throughout pregnancy. It is therefore imperative that we elucidate the mechanisms involved in regulating preimplantation embryonic responses to stress and that govern development. The MAPK pathways are involved in both responding to environmental stress and regulation of development throughout embryogenesis, and are therefore good candidates to study the mechanisms involved in preimplantation embryonic adaptation to stress and development. The preimplantation embryo culminates in the development of a fluid filled structure called the blastocyst. It is at this stage the first differentiation events occur and the trophectoderm (TE), which will go on to form the embryonic portion of the placenta, develops. The p38 MAPK is required for embryo development to proceed beyond the 8-16 cell stage as well to play an adaptive role in regulating embryonic response to culture stress. My hypothesis is that the MAPK
In a mouse model, in vitro fertilization or extended embryo culture leads to the increased expression of TRP53 in susceptible embryos. Ablation of the TRP53 gene improved embryo viability indicating that increased expression of TRP53 is a cause of the reduction of embryo viability resulting from in vitro fertilization or embryo culture. This study investigates the status of TRP53 expression in human embryos produced by intracytoplasmic sperm injection. Following fertilization, embryos were cultured for 96 h and then cryopreserved. Immediately upon thawing they were fixed in formaldehyde and subjected to immunostaining for TRP53. Staining was visualized by confocal microscopy. Negative controls were incubated with isotype control immunoglobulin and showed negligible staining. All embryos showed TRP53 staining above negative controls. TRP53 staining was heterogenous within and between embryos. An embryo that showed retarded development showed high levels of TRP53 expression. A blastocyst that had ...
A preparation and a method of making composite blastocysts (CBs) from aggregates of dissociated cells of non-viable pre-embryos are disclosed. The CB is characterized morphologically by having two distinct tissue types, the inner cell mass (ICM) and the trophectoderm (TE), and a blastocoelic cavity (BC). The ICM is differentially stainable with bisbenzimide and the TE is differentially stainable with propidium iodide. The ICM is pluripotent in that it contains embryonic stem (ES) cells. The TE cells are pluripotent in that they can give rise to all cell types normally derived from TE cells. The primate TE is characterized by the production of chorionic gonadotrophin. The method of making CBs is an aggregation process (AP) comprising inter alia the following steps: 1) dissociation of discarded pre-embryos; 2) isolation of single nucleated cells from dissociated discarded pre-embryos; 3) microsurgical encapsulation of several cells within a host zona pellucida or artificial aggregation with or without a
Bilaminar blastocyst or Bilaminar disc refers to the epiblast and the hypoblast, evolved from the embryoblast. These two layers are sandwiched between two balloons: the primitive yolk sac and the amniotic cavity. The inner cell mass, the embryoblast, begins to transform into two distinct epithelial layers just before implantation occurs. The epiblast is the outer layer that consists of columnar cells.The inner layer is called the hypoblast, or primitive endoderm, which is composed of cuboidal cells. As the two layers become evident, a basement membrane presents itself between the layers. The final two layers of the embryoblast are known collectively as the bilaminar embryonic disc as well as the bilaminar blastocyst or bilaminar blastoderm. This bilaminar blastocyst also defines the primitive dorsal ventral axis. Blastocyst implantation will occur during the second week of fetal development in the endometrium of the uterus; the epiblast is dorsal and the hypoblast is ventral. The zygote ...
The effect of glucose and insulin on the in vitro development of the rat preimplantation embryo was studied by incubating rat blastocysts recovered on days 5 or 6 of pregnancy in the absence or presence of increasing levels of glucose and/or insulin for 24 or 48 h. A differential cell-staining method allowed the separate counting of inner cell mass (ICM) and trophectoderm (TE) cells at the end of the incubation period. In a high-glucose medium (17 mM), ICM and, to a lesser extent, TE developments were significantly and irreversibly inhibited. Low insulin concentrations (3 pM) stimulated ICM and TE development in the presence of 1.1 or 6 mM glucose. Higher insulin levels (30-600 pM) in a 6-mM glucose medium, resulted in a dose-dependent inhibition of ICM and, to a lesser extent, TE development after both 24 and 48 h. This insulin-induced inhibition was reversible if insulin was removed from the medium after 24 h. In the absence of glucose in the medium, insulin was neither stimulatory nor ...
ICR albino mouse embryos (n=4lS0) were used to determine production of progesterone and estradiol. In Experiment I, cultures containing 20 (n=lO), 40 (n=lO) or 60 (n=6) early blastocysts were incubated in 13 X 100 rom tubes with .25 ml BMOC-2 for 20 h under 5% CO2 and air at 37C. Also, 20 (n=4) , 40 (n=7) and 60 (n=6) control embryos were frozen at -90C after flushing. Viability was determined by culturing 20 (n=5) , 40 (n=7) and 60 (n=6) for 24 h at which time percent normal development was microscopically evaluated.. In Experiment II, 40 embryos at either morula, early blastocyst or late blastocyst (n=lO) stage were cultured similarly. Viability and control steroid levels were determined on n=5, n=7 and n=7 cultures. Incubated and control cultures were extracted with diethyl ether and progesterone and estradiol isolated on Sephadex LH-20 columns prior to quantification by radioimmunoassay.. Viability for all cultures was 95.6 + .05% (~+SD). In Experiment I, incubated progesterone and estradiol ...
In the present study, we report that suppression of Nek2 expression by RNAi resulted in developmental defects at the second mitosis of the mouse early embryos. Many blastomeres in the Nek2-suppressed embryos were arrested at M phase with abnormal spindle structures. These results confirm that Nek2 is essential for embryonic mitosis, especially for proper segregation of chromosomes.. Phenotypes of Nek2-suppressed mouse embryos are comparable to those of the Xenopus embryos in which the Nek2 activity was reduced by microinjection of the mRNA for kinase-inactive form of Nek2B or of the Nek2 antibody (Uto and Sagata, 2000). Depletion of Nek2 resulted in fragmentation or dispersal of the centrosomes and eventually in interference of embryonic development of both species. In addition, our results are consistent with a previous report in which removal of the Nek2 proteins from Xenopus egg extracts did not disturb entry into mitosis and accompanying condensation of chromosomes (Fry et al., 2000). ...
Principal Investigator:SAIJO YASUO, Project Period (FY):2015-04-01 - 2017-03-31, Research Category:Grant-in-Aid for Challenging Exploratory Research, Research Field:Respiratory organ internal medicine
STUDY QUESTION: To what extent do patient- and treatment-related factors explain the variation in morphokinetic parameters proposed as embryo viability markers?. SUMMARY ANSWER: Up to 31% of the observed variation in timing of embryo development can be explained by embryo origin, but no single factor elicits a systematic influence.. WHAT IS KNOWN ALREADY: Several studies report that culture conditions, patient characteristics and treatment influence timing of embryo development, which have promoted the perception that each clinic must develop individual models. Most of the studies have, however, treated embryos from one patient as independent observations, and only very few studies that evaluate the influence from patient- and treatment-related factors on timing of development or time-lapse parameters as predictors of viability have controlled for confounding, which implies a high risk of overestimating the statistical significance of potential correlations.. STUDY DESIGN, SIZE, DURATION: ...
A.P. Wong et al, Directed differentiation of human pluripotent stem cells into mature airway epithelia expressing functional CFTR protein, Nat. Biotechnol., vol. 30, no. 9, pp. 876-882, Sept. 2012.. Y. Yamanaka et al, FGF signal-dependent segregation of primitive endoderm and epiblast in the mouse blastocyst, Development, vol. 137, no. 5, pp. 715-724, March 2010.. C. Chazaud et al, Early lineage segregation between epiblast and primitive endoderm in mouse blastocysts through the Grb2-Mapk pathway, Dev. Cell, vol. 10, no. 5, pp. 615-624, May 2006.. H. Niwa et al, Interaction between Oct3/4 and Cdx2 Determines trophectoderm differentiation, Cell, vol. 123, no. 5, pp. 917-929, Dec. 2005.. A. Nagy et al, Derivation of completely cell culture-derived mice from early-passage embryonic stem cells, Proc. Natl. Acad. Sci. U.S.A., vol. 90, no. 18, pp. 8424-8428, Sept. 1993. ...
Embryonic stem (ES) cells are derived from blastocyst-stage embryos and are thought to be functionally equivalent to the inner cell mass, which lacks the ability to produce all extraembryonic tissues. Here we identify a rare transient cell population within mouse ES and induced pluripotent stem (iPS) cell cultures that expresses high levels of transcripts found in two-cell (2C) embryos in which the blastomeres are totipotent. We genetically tagged these 2C-like ES cells and show that they lack the inner cell mass pluripotency proteins Oct4 (also known as Pou5f1), Sox2 and Nanog, and have acquired the ability to contribute to both embryonic and extraembryonic tissues. We show that nearly all ES cells cycle in and out of this privileged state, which is partially controlled by histone-modifying enzymes. Transcriptome sequencing and bioinformatic analyses showed that many 2C transcripts are initiated from long terminal repeats derived from endogenous retroviruses, suggesting this foreign sequence ...
article{1048354, abstract = {Recent studies have shown that short-term exposure of oocytes to a stressor such as hydrostatic pressure or osmotic stress might induce stress tolerance in embryos. The aim of the present study was to investigate the consequences of short-term hydrogen peroxide (H2O2) exposure to bovine in vitro matured cumulus-oocyte complexes (COCs) on subsequent preimplantation embryo development and apoptosis. in the first experiment, mature COCs were incubated in H2O2 at concentrations ranging between 0.01 and 100 mu mol/l, and subsequently fertilized and cultured. Oocyte incubation with 50-100 mu mol/l of H2O2 resulted in a significantly higher blastocyst yield (47.3\%) in comparison with control medium (31.8\%), while apoptotic cell ratio was inversely related with H2O2 concentration. In the second experiment, we showed that the stress tolerance after H2O2 exposure was not mediated by increased glutathione content in treated oocytes nor by enhanced fertilization or ...
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TY - JOUR. T1 - The in vitro development of blastocyst-derived embryonic stem cell lines. T2 - Formation of visceral yolk sac, blood islands and myocardium. AU - Doetschman, T. C.. AU - Eistetter, H.. AU - Katz, M.. PY - 1985/1/1. Y1 - 1985/1/1. N2 - The in vitro developmental potential of mouse blastocyst-derived embryonic stem cell lines has been investigated. From 3 to 8 days of suspension cultured the cells form complex embryoid bodies with endoderm, basal lamina, mesoderm and ectoderm. Many are morphologically similar to embryos of the 6- to 8-day egg-cylinder stage. From 8 to 10 days of culture about half of the embryoid bodies expand into large cystic structures containing alphafoetoprotein and transferrin, thus being analogous to the visceral yolk sac of the postimplantation embryo. Approximately one third of the cystic embryoid bodies develop myocardium and when cultured in the presence of human cord serum, 30% develop blood islands, thereby exhibiting a high level of organized ...
The inßuence of the sperm motility stimulant pentoxifylline (PF) on preimplantation embryo development in hamsters was evaluated. Eight-cell embryos were cultured in hamster embryo culture medium (HECM)-2, with or without PF (0· 0233·6 mM). There was 90%, 37% and 29% inhibition of blastocyst development by 3·6 (used for human sperm), 0·9 and 0 ·45 mM PF, respectively. However, 23 µM PF (exposed to hamster oocytes during IVF) signicantly (P , 0·05) improved blastocyst development (63· 6% v. 51· 8%); morulae development was, however, not curtailed by 0·45 mM or 0·9 mM PF (51·8%±6·0 or 50·5%±11·3, respectively). Post-implantation viability of PF-treated embryos was assessed by embryo transfer; 43% of 80 PF-treated embryos implanted compared with 40% of 79 control embryos. Of the 9 recipients, 6 females delivered pups (19, i.e. 16% of transferred embryos or 53% of implanted embryos). These data show that in hamsters, continuous presence of PF at 0·45-3·6 mM is detrimental to ...
LINE-1 (Long Interspersed Nuclear elements) and HERVs (Human Endogenous Retroviruses) are two families of retrotransposons which together account for about 28% of the human genome. Genes harbored within LINE-1 and HERV retrotransposons, particularly that encoding the reverse transcriptase (RT) enzyme, are generally expressed at low levels in differentiated cells, but their expression is up-regulated in embryonic tissues and transformed cells. Here we review evidence indicating that the LINE-1-encoded RT plays regulatory roles in early embryonic development. Indeed, antisense-mediated inhibition of expression of a highly expressed LINE-1 family in mouse zygotes caused developmental arrest at the two- or four-cell embryo stages. Development is also arrested when the embryo endogenous RT activity is pharmacologically inhibited by nevirapine, an RT inhibitor currently employed in AIDS treatment. The arrest of embryonic development is irreversible even after RT inhibition is removed and it is associated with
Principal Investigator:SUGIYAMA Fumihiro, Project Period (FY):1995 - 1996, Research Category:Grant-in-Aid for Scientific Research (C), Section:試験, Research Field:Laboratory animal science
Transcription factor control of TE/ICM segregation. TE and ICM lineage segregation is controlled by a small group of transcription factors. Specifically, Cdx2 is required for TE development, while the pluripotency markers octamer 3/4 (Oct4), Nanog, and SRY-box containing gene 2 (Sox2) are involved in establishing the ICM fate. In the mouse, Cdx2 is expressed at varying levels in all blastomeres starting at the eight-cell stage, but it becomes restricted to outside, future TE cells, prior to blastocyst formation (Figure 1) (72, 73). This variation in Cdx2 levels between individual blastomeres at the eight-cell stage may be a result of differences in the order and orientation of the cleavage divisions leading up to this stage (71). Embryos missing Cdx2 do form blastocysts initially, but the TE in these embryos loses its epithelial integrity and cannot differentiate further, resulting in death around the time of implantation (74). Oct4, Nanog, and Sox2 have expression patterns that are ...
Glutathione (C10H17N3O6S) is a tripeptide (γ-Glu-Cys-Gly) widespread in living organism. Glutathione (GSH) at the 5 mM concentration increased the motility and fertility of frozen-thawed sperm. Intracellulair glutathione improved the cleavage rate and embryo development to the blastocyst rate. Research on in vitro embryos production through the implementation of GSH during IVF (in vitro fertilization) on embryo development has been conducted at the Laboratorium Reproductive of Physiology, Research Institute for Animal Production. Ovaries of beef cattle as source of oocytes were collected from the slaughterhouse in a thermo flask with 350C PBS as medium and transported to the laboratory. The oocytes were fertilized in vitro with selected motile sperm using Percoll gradient (90 and 45%). Ten COCs (cumulus oocytes complexes) were transfered to 44 μl of fertilization medium (mTALP) was performed with either 0; 0.25; 0.50; 0.75 and 1.00 mM of glutathione as treatment A, B, C, D and E respectively, ...
More than 90 percent of enucleated one-cell mouse embryos receiving pronuclei from other one-cell embryos successfully develop to the blastocyst stage in vitro. In this investigation, nuclei from successive preimplantation cleavage stages were introduced into enucleated one-cell embryos and the embryos were tested for development in vitro. Although two-cell nuclei supported development to the morula or blastocyst stage, four-cell, eight-cell, and inner cell mass cell nuclei did not. The inability of cell nuclei from these stages to support development reflects rapid loss of totipotency of the transferred nucleus and is not the result of simultaneous transfer of membrane or cytoplasm.
She said that her results also had implications for human embryo stem (ES) cell research. Human embryonic stem cell lines are derived from blastocysts that, we know now, already have, or still have, a form of XCI. While mouse embryos reactivate the X chromosome in the inner cell mass at the blastocyst stage so that the derived embryonic stem cells are completely undifferentiated, it is not yet known whether this occurs in human embryos. The onset and subsequent steps of XCI in human pre-implantation embryos occur at a later stage than in mouse embryos. Thus, it is possible that reactivation of the X chromosome happens also at a later stage, after the usual time for ES cell derivation. The current human ES cell lines may, therefore, still have the first wave of XCI. Indeed, the majority of human ES cells have XCI features ...
Blastocyst transfer is associated with a higher chance for pregnancy. The risk for multiple pregnancy can be reduced by transferring fewer blastocysts.
The precise relationship of embryonic stem cells (ESCs) to cells in the mouse embryo remains controversial. We present transcriptional and functional data to identify the embryonic counterpart of ESCs. Marker profiling shows that ESCs are distinct from early inner cell mass (ICM) and closely resemble pre-implantation epiblast. A characteristic feature of mouse ESCs is propagation without ERK signalling. Single-cell culture reveals that cell-autonomous capacity to thrive when the ERK pathway is inhibited arises late during blastocyst development and is lost after implantation. The frequency of deriving clonal ESC lines suggests that all E4.5 epiblast cells can become ESCs. We further show that ICM cells from early blastocysts can progress to ERK independence if provided with a specific laminin substrate. These findings suggest that formation of the epiblast coincides with competence for ERK-independent self-renewal in vitro and consequent propagation as ESC lines. It has been unclear at which stage of
At Atlantic Reproductives embryology lab, all embryos are cultured to the blastocyst development stage, which is the best stage for doing genetic analysis.
The ultrastructure of bovine morulae and blastocysts developed from in vitro-matured and -fertilized oocytes in a serum-supplemented medium was compared with that of morulae and blastocysts collected
Blastocyst culture is for improved take home baby rate. But blastocyst culture may reduce the number of embryo transfers as all embryos may not grow to the blastocyst stage & may be arrested at 4 or 6 cell stage, thus resulting in cancellation of the entire cycle. Blastocyst culture. ...
To investigate the relationship between air bubble position after blastocyst transfer (BT) and pregnancy rates (PRs). Retrospective cohort study. University-based infertility center. Three hundred fifteen consecutive nondonor BTs by
1. Li X-Y, Thomas S, Sabo PJ, Eisen MB, Stamatoyannopoulos JA, Biggin MD. The role of chromatin accessibility in directing the widespread, overlapping patterns of Drosophila transcription factor binding. Genome Biol. 2011;12: R34. doi: 10.1186/gb-2011-12-4-r34 21473766. 2. Thurman RE, Rynes E, Humbert R, Vierstra J, Maurano MT, Haugen E, et al. The accessible chromatin landscape of the human genome. Nature. 2012;489: 75-82. doi: 10.1038/nature11232 22955617. 3. Klemm SL, Shipony Z, Greenleaf WJ. Chromatin accessibility and the regulatory epigenome. Nat Rev Genet. 2019;20: 207-220. doi: 10.1038/s41576-018-0089-8 30675018. 4. Wu J, Huang B, Chen H, Yin Q, Liu Y, Xiang Y, et al. The landscape of accessible chromatin in mammalian preimplantation embryos. Nature. 2016;534: 652-657. doi: 10.1038/nature18606 27309802. 5. Clark SJ, Argelaguet R, Kapourani C-A, Stubbs TM, Lee HJ, Alda-Catalinas C, et al. scNMT-seq enables joint profiling of chromatin accessibility DNA methylation and transcription in ...
Embryonic stem cells derived from the inner cell mass of blastocyst-stage embryos are totipotent cells that can differentiate into all tissues and cell types (1). Recent discoveries that extend the potential use of ES cells include the isolation of ES cells from embryonic human tissue (2) and transplantation in sheep and mice of nuclei from mature tissues into enucleated oocytes, permitting the generation of ES cells from the same individual (3). Thus, ES cell technology may be the basis of new cell replacement therapies.. ES cells induced to differentiate in vitro give rise to many cell types including hematopoietic precursors, heart and skeletal muscle, endothelium, and neural cells (4). In the central nervous system, proliferation and differentiation of multipotential neural stem cells and glial progenitors can be controlled by defined factors (5). Here, we show that applying this knowledge to ES cells permits efficient in vitro generation of precursors for oligodendrocytes and astrocytes. ...
Stress in utero or during the early post natal period plays a cardinal role in shaping future health. An initial stressor, occurring when the embryo is exquisitely sensitive to environmental changes, reprograms the developing gene networks, tissue, and organs so that the resulting individual is predisposed to disease. We hold that suboptimal preimplantation conditions predispose adult individuals to altered glucose homeostasis, leading to an increased incidence of insulin resistance. Our preliminary data show that increased stress during in vitro culture results in proportionate deterioration of gene expression in mouse embryos. Most importantly, we have identified thioredoxin-interacting protein (txnip) as a marker of preimplantation stress. Txnip mRNA and protein levels are upregulated in blastocyst after in vitro culture; suboptimal culture conditions (Whittens medium- WM) results in a more intense txnip upregulation than culture in optimized medium (KSOM with amino acid, KAA). Txnip ...
Embryonal carcinoma (EC) cells have served as a model to study the relationship between cancer and cellular differentiation given their potential to produce tumors and, to varying degrees, participate in embryonic development. Here, nuclear transplantation was used to assess the extent to which the tumorigenic and developmental potential of EC cells is governed by epigenetic as opposed to genetic alterations. Nuclei from three independent mouse EC cell lines (F9, P19, and METT-1) with differing developmental and tumorigenic potentials all were able to direct early embryo development, producing morphologically normal blastocysts that gave rise to nuclear transfer (NT)-derived embryonic stem (ES) cell lines at a high efficiency. However, when tested for tumor or chimera formation, the resulting NT ES cells displayed an identical potential as their respective donor EC cells, in stark contrast to previously reported NT ES cells derived from transfer of untransformed cells. Consi stent with this ...
Human embryonic stem cells are pluripotent cells produced from the internal cell mass of preimplantation stage embryos. genes like the human being thyroid transcription element 1 (and also have overlapping temporal and spatial expressions in the peripheral epithelial cells from the developing lung where activates the transcription of (Shaw-White manifestation is directly controlled through this synergistic actions from the N-terminal and zinc-finger domains of as well as the homeodomain area of (Liu in mouse embryonic stem (mES) cells offers been proven to induce differentiation towards extraembryonic endoderm a prerequisite for lung organogenesis (Fujikura (SRY (sex-determining area Y) package 17) a marker of definitive endoderm in mice offers revealed the key function of the element in the differentiation of respiratory epithelial cells into the various cells of the conducting airways (Park when grown in suspension and form embryoid bodies (EBs) which express markers specific to the three ...
Researchers supported the use of the portal vein as a site for islet transplantation, but noted that there are issues with injecting encapsulated cells into the liver. Finally, the research team suggested decreasing capsule size to nano-scale and combining PEGylation coating of capsules with a layer of poly(ethylene) glycol molecules with low-dose immunosuppression to improve engraftment and long-term function.. What about stem cells?. Human embryonic stem cells emerge five to seven days into an embryos development, when the embryo is a hollow sphere. The sphere consists of an outer layer of cells that goes on to form the placenta, and an inner cluster of cells known as the inner cell mass that goes on to form all of the tissues of the body. At this stage, the embryo is known as a blastocyst. Embryonic stem cells arise from the inner cell mass. (Picture shows a five-day-old embryo, known as a blastocyst. The dark patch inside the blastocyst is the inner cell mass, from which embryonic stem ...
ObjectiveTwo critical points of early development are the first and second lineage segregations, which are regulated by a wide spectrum of molecular and cellular factors. Gene regulatory networks, are one of the important components which handle inner cell mass (ICM) and trophectoderm (TE) fates and the pluripotency status across different mammalian species. Considering the importance of goats in agriculture and biotechnology, this study set out to investigate the dynamics of expression of the core pluripotency markers at the mRNA and protein levels.Materials and MethodsIn this experimental study, the expression pattern of three pluripotency markers (Oct4, Nanog and Sox2) and the linage specific markers (Rex1, Gata4 and Cdx2) were quantitatively assessed in in vitro matured (MII) oocytes and embryos at three distinctive stages: 8-16 cell stage, day-7 (D7) blastocysts and D14 blastocysts. Moreover, expression of Nanog, Oct4, Sox2 proteins, and their localization in the goat blastocyst was observed
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Both pluripotent Embryonic Come Cells (ESCs), established from preimplantation murine blastocysts, and Epiblast Come cells (EpiSCs), established from postimplantation embryos, can self-renew in culture or differentiate into each of the primary germ layers. and their self-renewal requires Activin and FGF2. While the primary TFs April4, SOX2, and NANOG are indicated in both pluripotent cell types, ESCs and EpiSCs screen unique gene appearance users, and many extra TFs Zaurategrast that are essential for ESC self-renewal are Zaurategrast lacking in EpiSCs [4, 6]. Therefore ESCs and EpiSCs possess been posited to represent two unique claims highlighting the developing growth phases of the epiblast and and and was equal in both pluripotent cell types although appearance was somewhat downregulated in EpiSCs. These microarray data had been authenticated for a subset of genetics using qRT-PCR of mRNA singled out from our ESCs and EpiSCs (Helping Details Fig. T4). We after that analyzed the FAIRE ...
TY - JOUR. T1 - Proliferation-stimulating effect of colony stimulating factor 2 on porcine trophectoderm cells is mediated by activation of phosphatidylinositol 3-kinase and extracellular signal-regulated kinase 1/2 mitogen-activated protein kinase. AU - Jeong, Wooyoung. AU - Kim, Jinyoung. AU - Bazer, Fuller W.. AU - Song, Gwonhwa. N1 - Copyright: Copyright 2014 Elsevier B.V., All rights reserved.. PY - 2014/2/10. Y1 - 2014/2/10. N2 - Colony-stimulating factor 2 (CSF2), also known as granulocyte macrophage colony-stimulating factor, facilitates mammalian embryonic development and implantation. However, biological functions and regulatory mechanisms of action of porcine endometrial CSF2 in peri-implantation events have not been elucidated. The aim of present study was to determine changes in cellular activities induced by CSFs and to access CSF2-induced intracellular signaling in porcine primary trophectoderm (pTr) cells. Differences in expression of CSF2 mRNA in endometrium from cyclic and ...
Cell division. During the first 12 hours after conception, the fertilized egg cell remains a single cell. After approximately 30 hours, it divides from 1 cell into 2 and 15 hours later, the 2 cells divide into 4. And at the end of 3 days, the fertilized egg cell has become a berry-like structure made up of 16 cells. This structure is called a morula, which is Latin for mulberry. The cells continue to divide 8 or 9 days following conception into a blastocyst. Although it is only the size of a pinhead, the blastocyst is composed of hundreds of cells. The blastocyst is slowly carried by tiny hair-like projections in the fallopian tube called cilia toward the uterus. During the critically important process of implantation, it must attach itself to the uterine lining where it will be able to get nourishment from the mothers blood supply. If the blastocyst is unable to attach, the pregnancy will fail to survive. ...
MELO, I.M.F. et al. Effect of enrofloxacin on the blastocyst endometrial interactions and their impact on placental and fetal development in rats. Arq. Bras. Med. Vet. Zootec. [online]. 2014, vol.66, n.5, pp.1406-1412. ISSN 0102-0935. http://dx.doi.org/10.1590/1678-5594.. Some studies have shown the toxic effects of enrofloxacin in various tissues. Thus, the hypothesis that enrofloxacin could interfere with placental development and generate adverse effects to the fetus was tested in this study. Enrofloxacin (Baytril(r)) was administered in the dose of 5mg/kg daily, i.m., throughout gestation in rats. The placentas were analyzed morphologically, morphometrically, and immunohistochemically on the 7, 14, and 20th days of pregnancy. The results showed that enrofloxacin reduced the number of implantation sites, weight, and placental disk total area at 14 and 20 days of development, in addition to the element components of the placenta. The histochemical analysis did not reveal significant changes in ...
Developed in the early 1990s, PGD is used so couples can prevent a pregnancy affected by a genetic condition or chromosomal disorder. Its performed by removing one or two cells (called blastomeres) for biopsy from the preimplantation embryo at the six to ten cell stage (about day three of development). If one or both parents-to-be have a known genetic abnormality and their child might be at increased risk for Tay Sachs disease, cystic fibrosis, muscular dystrophy, Fragile X syndrome, spinal muscular atrophy or other conditions, PGD can show if the embryo is likely to grow into a person with that potential problem. If thats the case, most like a decision will be made not to implant a specific embryo in a womans womb.. While its almost hard to believe, no rigorous long-term studies have been carried out in order to see whether PGD poses any serious health risks down the line - even though the procedure involves manipulating a developing embryo. So Chinese scientists Ran Huo, Qi Zhou and ...
The number of supernumerary vitrified blastocysts correlates positively with the odds of implantation and live birth in single-blastocyst transfers.
We performed in vitro maturation (IVM) and in vitro fertilization (IVF) of the d3 PGCLC- and PGC-derived oocytes and the WT oocytes at 3 weeks (fig. S6B). Despite differences in COC stability and shape, the PGCLC-derived oocytes reached metaphase II (MII), were fertilized, and developed into two-cell embryos with an efficiency comparable to that of oocytes from other sources (Fig. 2B, fig. S6C, and Table 1). Some of the two-cell embryos from the PGCLCs developed further into blastocysts in vitro [19 of 46 (19/46), ~39%] (Fig. 2B). We transferred the two-cell embryos from PGCLCs, as well as those from the other sources, to separate foster mothers. We obtained newborn pups from the two-cell embryos derived from PGCLCs (5/127, ~3.9%), as well as from those derived from E12.5 PGCs (13/75, ~17.3%) and WT 3-week oocytes (7/55, ~12.7%) (Fig. 2C, fig. S7A, and Table 1). All of these offspring grew similarly into adulthood (fig. S7C). The PGCLC-derived offspring bore the BVSC transgenes, a normal ...
The possibility of producing transgenic buffalo embryos by chimera and nuclear transfer (NT) using buffalo embryonic germ (EG)-like cells expressing enhanced green fluorescent protein (EGFP) has been explored in this study. Buffalo EG-like cells and fibroblasts with two to eight passages were transfected with the lined plasmid (pCE-EGFP-IRES-Neo-dNdB) using LipofectamineTM 2000 and selected by culturing in 200 μg/ml G418 for 6-8 days. G418 resistant fibroblasts and EG-like cells were used for embryo chimera and NT. To produce blastocysts by chimera, 8-16 cells embryos were injected with EG-like and fibroblast cells. Then, to produce blastocysts by NT, in vitro maturated oocytes were enucleated and afterwards EG-like/fibroblast cells transferred into the perivitelline space. No statistical differences were observed for the total blastocyst produced by the chimeric method, using EG-like and fibroblasts as donor cells, resulting on an accomplishment of 35.6% vs 33.3%, respectively. Nevertheless, ...
Hi, My Dr. did transfer of four blastocysts on day 7 and I am wondering if there are any success stories about transfering this late.
When scientists discovered how to reprogram skin cells into neurons, research on neurological disorders broadened. Disorders of the brain have been historically difficult to study because brain tissue cannot be taken from a living person; human brain tissue can only be collected post-mortem.1 Following the discovery of induced pluripotent stem cell (iPSC) technology, skin samples from patients with neurological disorders could be reprogrammed into neurons and used to observe how living neurons are altered in disease states.2 While reprogrammed human skin cells have provided novel models of neurological disorders, the approach has always been limited because neurons are grown in a dish, a markedly different environment from how they exist within the body.1. Chang and colleagues recently provided the groundwork to produce an improved model compared to neurons in a dish: the ability to recreate an entire portion of the brain in vivo.3 The blastocyst complementation technique was utilized in this ...
Fig. 4. ESR1 recruits HDAC1 to the ASE region to exclude p300 from the Xnr1-dependent transcriptional complex. (A) The experimental strategy of animal cap assay. Zygotic transcription starts at stage 8. (B) Twenty pg activin RNA was injected into 2-cell-stage embryos with or without 100 pg E1A RNA and animal caps were dissected at the stage 8. Caps were cultured until sibling embryos reached stage 10, and Pitx2 gene expression was evaluated by semi-quantitative RT-PCR analysis. (C) Two ng flag-p300 RNA and/or 20 pg activin RNA was injected into 2-cell-stage embryos with or without 2 ng HA-ESR1 RNA, and embryonic extracts were isolated at stage 10 for ChIP analysis. ChIP assays were performed using α-flag antibody. (D) Twenty pg activin RNA was injected into 2-cell-stage embryos with or without 2 ng HA-ESR1 RNA, and animal caps were dissected at the stage 8. Caps were cultured with or without 50 nM TSA until sibling embryos reached stage 10, and Pitx2 gene expression was evaluated by ...
This study demonstrated that bovine sperm exposure to GAGs positively affected sperm fertilizing ability, in vitro embryo developmental potential, and embryonic gene expression. This is the first report to determine concurrently the effects of four different GAGs (HP, HA, CD, and DS). HP was the most potent GAG for enhancing sperm motility and inducing the acrosome reaction. HP exposed sperm exhibited improved 2 PN formation, cleavage rate, blastocyst formation rate, and embryo cell numbers relative to the control (p,0.05). Additionally, in embryos developing from fertilization with HP-treated sperm significant changes in gene expression were detected in genes involved in pluripotency (Oct4, upregulated), apoptosis (Bax inhibitor, upregulated; Caspase 3, downregulated), and cell growth (Glut5, upregulated), relative to control embryo gene expression (p,0.05). Sperm exposure to HA resulted in intermediate levels of changes, and, similar to HP, HA treatment of sperm resulted in significantly ...
During late peri-implantation development, porcine conceptuses undergo a rapid (2C3 hrs) morphological transformation from a 10 mm sphere to a thin filamentous form greater than 150 mm in length. were collected from pregnant gilts and subjected to SSH. Forward and reverse subtractions 212701-97-8 supplier were performed to identify 212701-97-8 supplier candidate genes differentially indicated during … Continue reading During late peri-implantation development, porcine conceptuses undergo a rapid (2C3 hrs). ...
By 4 days after conception (well before implantation) the human zygote (what those who want abortion-on-demand misidentify as a fertilized egg) gives rise to the blastocyst stage. The blastocyst has 2 cell layers - the trophectoderm, which becomes the placenta, and the inner cell mass, which forms the fetus. The placenta is only partially the mothers, and is mostly developed by the baby. In fact, the tissues of the baby may touch those of the mother, but they do not merge. The blood supply of the mother is never in contact with that of the baby. I write about the placenta, because that is the only point where there could be contention on where the babys body ends and the womans body begins ...
J:170133 Kim ST, Marquard K, Stephens S, Louden E, Allsworth J, Moley KH, Adiponectin and adiponectin receptors in the mouse preimplantation embryo and uterus. Hum Reprod. 2011 Jan;26(1):82-95 ...
embryo hatching.. Standard IVF protocols include culturing of embryos within the laboratory for three days, followed by transfer of cleavage stage embryos (6 to 8 cells), on Day 3, to the uterine cavity. Following transfer, the embryos must continue to progress to the blastocyst stage, shed the ZP, and embed into the uterine wall. In 1989 Cohen and his co-investigators observed a higher implantation rate in patients undergoing IVF, who had the ZP of their embryos mechanically opened. They therefore hypothesized that artificially creating a gap in the ZP might serve to facilitate embryo hatching and implantation. Microscopic manipulation of the ZP, in order to augment hatching and implantation, subsequently became known as assisted hatching. Prospective randomized clinical studies have been performed in order to evaluate the effectiveness of assisted hatching. Several studies report a significant increase in embryo implantation and clinical pregnancy rates, in select groups of patients whose ...
Preimplantation genetic testing or preimplantation genetic diagnosis is a technique in which the embryos prepared through in vitro fertilization are tested for defects before implantation. Preimplantation genetic testing enables physicians and to identify the defects present in the embryos and selectively implant he...
Although the sheep has been the first animal to be cloned by somatic cells nucleartransfer (SCNT), post natal survival of cloned lambs seems to be lower than the otheranimal cloned so far. We present the results of a large scale cloning experiment designed toexplore the causes of peri- and post-natal mortality of cloned lambs. Blastocysts (n. 93)obtained by nuclear transfer of somatic cells (granulosa cells), were transferred into 41recipient ewes, and pregnancies were monitored by ultrasound scanning. In vitro derived,fertilized embryos (IVF, n=123) were also transferred as a control. Pre and early postimplantation development was comparable between cloned and control IVF embryos;however, dramatic losses occurred from the first month of pregnancy onward, with only 12out of 93 (13%) clones reaching full-term development, compared to 51 out of 123 (41.6%)lambs born from the IVF control embryos. Blood samples collected from the cloned lambsafter birth, revealed a wide range of abnormalities ...
To be able to perform implantation on the uterine wall, the blastocyst first needs to get rid of the zona pellucida. This lysis ... Prior to this event, the predecessor of the embryo, in the form of a blastocyst, is surrounded by a glycoprotein sphere called ... In some situations, the term "hatching" is used only for artificial ways to free the blastocyst from the zona pellucida, and ... "Blastocyst Development". UNSW Embryology. UNSW CRICOS. ISBN 978 0 7334 2609 4. Langman's Medical Embryology 6th Edition, page ...
This results in a hollow ball of cells known as the blastocyst. The blastocyst's outer cells will become the first embryonic ... A morula is distinct from a blastocyst in that a morula (3-4 days post fertilization) is a 16-cell mass in a spherical shape ... "The Morula and Blastocyst". the Endowment for Human Development. Retrieved 11 April 2015. Sherman, Lawrence S. et al., eds. ( ... A morula, if untouched and allowed to remain implanted, will eventually develop into a blastocyst. The morula is produced by a ...
At this stage, the embryo is called a blastocyst. Embryogenesis Blastocyst Forgács, G.; Newman, Stuart A. (2005). "Cleavage and ...
Nevertheless, the apposition on the blastocyst is not dependent on if it is on the same side of the blastocyst as the inner ... There is massive communication between the blastocyst and the endometrium at this stage. The blastocyst signals to the ... perhaps because it increases the area of contact with the rather spherical blastocyst. On the blastocyst, on the other hand, it ... Factors from the blastocyst also trigger the final formation of decidual cells into their proper form. In contrast, some ...
However, if the structural integrity of the blastocyst is compromised prior to the experiment, the ICM is susceptible to the ... This technique is used to isolate the inner cell mass of the blastocyst. The trophoectoderm's cell junctions and tight ... Cruz, Y. P.; Treichel, R. S.; Harsay, E.; Chi, K. D. (1993-01-01). "Mouse Blastocyst Immunosurgery with Commercial Antiserum to ... Immunosurgery is a method of selectively removing the external cell layer (trophoblast) of a blastocyst through a cytotoxicity ...
spelling blastocoele, blastocele) is also termed the blastocyst cavity (or cleavage or segmentation cavity) is the name given ... Borland, Raymond Michael (1977). "Transport processes in the mammalian blastocyst". Development in Mammals. 1: 31-67. Cherr, ... called a blastocyst. for sex call 7838725002. Similar to mammals, fertilization of the avian ovum occurs in the oviduct. From ... to the fluid-filled cavity of the blastula (blastocyst) that results from cleavage of the oocyte (ovum) after fertilization. It ...
A hollow cavity forms marking the blastocyst stage. (day 1.5-3 of fertilization.) The blastocyst contains only a thin rim of ... The blastocyst reaches the uterus at roughly the fifth day after fertilization. It is here that lysis of the zona pellucida ... The blastocyst is fully implanted day 7-12 of fertilization. Formation of the yolk sac. The embryonic cells flatten into a disk ... This stage is called a blastocyst. Up to this point there is no growth in the overall size of the embryo, as it is confined ...
Clemetson CA, Moshfeghi MM, Mallikarjuneswara VR (1970). "Electrophoretic mobility of the rat blastocyst". Contraception. 1: ...
Blastocoel Blastocyst Oocyte Blastomere Encyclopædia Britannica. Encyclopædia Britannica Online. Encyclopædia Britannica Inc., ...
In mammals the blastula is referred to as a blastocyst. The blastocyst contains an embryoblast (or inner cell mass) that will ... In the mammalian blastocyst (term for mammalian blastula) there are three lineages that give rise to later tissue development. ... ISBN 978-0-7923-8336-9. Forgács & Newman, 2005: p. 27 Cockburn, Katie; Rossant, Janet (1 April 2010). "Making the blastocyst: ... Blastocyst Cellular differentiation Gastrulation Polarity in embryogenesis Diploblasty Triploblasty "Blastula". Encyclopædia ...
The hybrid cell is then stimulated to divide by an electric shock, and when it develops into a blastocyst it is implanted in ... Albieri then implanted the blastocyst into Dora's ovary. At this time, Dora does not know that she was carrying a clone. When ...
Early blastocyst stage 4 Random X-inactivation in the embryonic lineage (inner cell mass) Late blastocyst stage Late blastocyst ... XaP XaM → undergoing random X-inactivation in the embryonic lineage (inner cell mass) in the blastocyst stage, leading to: ... In the early blastocyst, this initial, imprinted X-inactivation is reversed in the cells of the inner cell mass (which give ... XiP XaM → undergoing X-activation in the early blastocyst stage, leading to: ...
Frankenberg, Stephen R.; De Barros, Flavia R.O.; Rossant, Janet; Renfree, Marilyn B. (2016). "The Mammalian Blastocyst". Wiley ...
Blastocyst Gravindex Wilcox AJ, Baird DD, Weinberg CR (1999). "Time of implantation of the conceptus and loss of pregnancy". ...
The blastocyst must not be confused with the blastula; even though they are similar in structure, their cells have different ... Mammals at this stage form a structure called the blastocyst,[1] characterized by an inner cell mass that is distinct from the ...
"Monozygotic twins and triplets in association with blastocyst transfer". Journal of Assisted Reproduction and Genetics. 21 (4 ...
hCG-positive indicates an implanted blastocyst and mammalian embryogenesis. These can be done to diagnose and monitor germ cell ...
At 148 hours, early blastocysts form. At 10-12 days, implantation occurs. The gestation period for cats is between 64 and 67 ...
As embryonic stem cells are derived from the inner cell mass at the blastocyst stage, removing them from the inner cell mass ... Stewart CL, Kaspar P, Brunet LJ, Bhatt H, Gadi I, Köntgen F, Abbondanzo SJ (September 1992). "Blastocyst implantation depends ...
A blastocyst is then formed and implanted in the uterus. Embryogenesis continues with the next stage of gastrulation, when the ... The decidua here is termed the decidua basalis; it lies between the blastocyst and the myometrium and forms the maternal part ... The epiblast is adjacent to the trophoblast and made of columnar cells; the hypoblast is closest to the blastocyst cavity and ... The resulting increase in size of the blastocyst causes it to hatch through the zona pellucida, which then disintegrates. The ...
Enders A. C., Lantz K. C., Schlafke S. (1990). Differentiation of the inner cell mass of the baboon blastocyst. Anat. Rec. 226 ... In mouse embryo, the visceral endoderm develop from the primitive endoderm of the blastocyst during the implantation stage ...
The success rate of ICSI is 23-44% blastocyst development. Domestication of the horse Endometrosis Evolution of the horse ... the egg might have already reached the blastocyst stage. The gestation period lasts for about eleven months, or about 340 days ...
At this point, the exocoelomic cavity replaces the blastocyst cavity. At days 11 to 12, there is further delineation of the ...
Reactivating tammar wallaby blastocysts oxidise glucose. Biology of Reproduction 58, 1425-1431. Spindler R.E., M.B. Renfree, G ... Reactivating tammar wallaby blastocysts oxidise fatty acids and amino acids. Journal of Reproduction and Fertility 115, 79-86. ... Oocyte metabolism predicts the development of cat embryos to blastocyst in vitro. Molecular Reproduction and Development 56, ... Carbohydrate uptake by quiescent and reactivating mouse blastocysts. Journal of Experimental Zoology 276, 132-137. Spindler R.E ...
Children born from vitrified blastocysts have significantly higher birthweight than those born from non-frozen blastocysts. ... "Birth after replacement of hatching blastocyst cryopreserved at expanded blastocyst stage". Lancet. 1: 647. PMID 2857991. CS1 ... when embryos are at the blastocyst stage, it can also be called elective single blastocyst transfer (eSBT). It significantly ... "Comparison of pregnancy outcomes in elective single blastocyst transfer versus double blastocyst transfer stratified by age". ...
Children born from vitrified blastocysts have significantly higher birthweight than those born from non-frozen blastocysts. For ... May 2010). "Obstetric outcomes after transfer of vitrified blastocysts". Hum Reprod. 25 (7): 1699-707. doi:10.1093/humrep/ ... from fertilisation to the blastocyst stage. Embryo cryopreservation is useful for leftover embryos after a cycle of in vitro ...
However, the egg can split into two, but still have one blastocyst. This will lead to one inner cell mass and one blastocyst. ... If the egg splits in the early blastocyst stage, two inner cell masses will be present, eventually leading to the twins sharing ...
Gardner, R.L. and Edwards, R.G. (1968). Control of the sex ratio at full term in the rabbit by transferring sexed blastocysts. ... Blastocyst injection was later adopted almost universally for assessing the developmental potential of embryonic stem (ES) ... Gardner, R. L.; Barton, S. C.; Surani, M. A. (1 October 1990). "Use of triple tissue blastocyst reconstitution to study the ... Gardner, R. L. (2007) The axis of polarity of the mouse blastocyst is specified before blastulation and independently of the ...
This new structure with a cavity in the center and the developing cells around it is known as a blastocyst. The presence of the ... Then, a flat layer cell forms on the exterior of this cavity, and the zona pellucida, the blastocyst's barrier, remains the ... The inner cell mass of the blastocyst divides rapidly, forming two layers. The top layer becomes the embryo, and cells from ... The cells on the exterior of the blastocyst begin excreting an enzyme which erodes epithelial uterine lining and creates a site ...
As a result, they decided to work with cultured blastocyst cells. The nuclear material of these blastocyst cells would be ...
Establishment of embryonic stem cells from rat blastocysts.. Kawamata M1, Ochiya T. ...
A blastocyst is a cellular mass that forms early in the embryo development process in mammals. Humans develop a blastocyst ... So a blastocyst is very early pregnancy? Is a blastocyst before implantation? It sounds like it would have to be if part of the ... Apparently, a blastocyst isnt an embryo yet. (In humans, the term embryo only applies after implantation.) At the blastocyst ... Within the blastocyst, there are two types of cells. In the interior is the inner cell mass, a portion of which will begin to ...
Waiting for blastocyst transfer! Hi everyone.. This is IVF #4 for us and were TTC #1. Its been a few years since we did IVF, ... We are hoping to get at least 2 blastocysts to transfer on day 5. I am so worried. I have previously only ever had day-2 ... Blastocyst transfer: How did your eggs go?. By Green Mum to be in forum Egg Donation ... Blastocyst transfer: How man embryos progressed?. By Green Mum to be in forum IVF ...
My transfer was 5 days ago using two Grade A blastocyst. They are from a donor, so I have not had any trigger injections or any ... 2dpt... Embryo is now a blastocyst 3dpt....Blastocyst hatches out of shell on this day 4dpt.. Blastocyst attaches to a site on ... 0dpt... Embryo is now a blastocyst 1dpt....Blastocyst hatches out of shell on this day 2dpt.. Blastocyst attaches to a site on ... 2dpt... Embryo is now a blastocyst 3dpt....Blastocyst hatches out of shell on this day 4dpt.. Blastocyst attaches to a site on ...
Generation Functional Lungs Conditional Blastocyst Complementation Using Pluripotent PubMed Journal Articles published on ... Building Blastocysts from Stem Cells.. The blastocyst stage and the subsequent implantation are critical for a successful ... A single mouse blastomere from an embryo until the 8-cell stage can generate an entire blastocyst. Whether laboratory-cultured ... 2019) describe a novel way to generate blastocyst-like structures only from pluripotent stem cells. These structures mimic ...
We are the leading providers with the highest record of Blastocyst culture & assisted hatching in England and Wales. Find out ... Blastocyst culture. Studies suggest that pregnancy rates improve if the embryos are left to develop to their blastocyst stage. ... Blastocyst Culture at the LWC. The London Womens Clinic now uses blastocyst culture as part of its standard laboratory ... Blastocyst Grading. Blastocyst grading or quality is determined by evaluating the outer ring of cells, known as the ...
A preparation and a method of making composite blastocysts (CBs) from aggregates of dissociated cells of non-viable pre-embryos ... One patent referring to blastocysts, U.S. Pat. No. 5,541,081, discusses the use of biochemical markers. The use of blastocysts ... Blastocysts typically develop on the fifth or sixth day of pre-embryo culture in vitro. At this time, they typically contain ... A blastocyst is composed of two tissue types 1) a distinct inner cell mass (ICM) placed eccentrically in the blastocoelic ...
The data suggest that insulin and glucose might interact and modulate blastocyst development as a function of their respective ... Stimulatory and Inhibitory Effects of Glucose and Insulin on Rat Blastocyst Development in Vitro. ... Stimulatory and Inhibitory Effects of Glucose and Insulin on Rat Blastocyst Development in Vitro ... Stimulatory and Inhibitory Effects of Glucose and Insulin on Rat Blastocyst Development in Vitro ...
Recent reports show that more fertility clinics in the country implement the procedure of blastocyst transfers to decrease the ... The blastocyst transfer or "Day5 Transfer" allows fertilized eggs to grow for an additional 2 days past the standard three-day ... Once the embryo reaches the blastocyst stage, the embryologist can better evaluate the quality of the embryo and to increase ... More Fertility Clinics Try Blastocyst Transfers. Posted on April 15th, 2003. Recent reports show that more fertility clinics in ...
"Deep learning as a predictive tool for fetal heart pregnancy following time-lapse incubation and blastocyst transfer, Human ... blastocysts with the potential of developing FH and accurately detect Tran D, Cooke S, Illingworth PJ, Gardner DK. Deep ... blastocysts with the potential of developing FH and accurately detect Tran D, Cooke S, Illingworth PJ, Gardner DK. Deep ... Deep learning as a predictive tool for fetal heart pregnancy following time-lapse incubation and blastocyst transfer. Chavez- ...
Organ culture as a new method for studying the implantation of mammalian blastocysts. T. W. Glenister ... Organ culture as a new method for studying the implantation of mammalian blastocysts ... Organ culture as a new method for studying the implantation of mammalian blastocysts. ... Organ culture as a new method for studying the implantation of mammalian blastocysts ...
Presentation] Generation of Lung Organ from Embryonic Stem cells via Blastocyst Complementation in Mice2017. *. Author(s). ... GENERATION OF LUNG ORGAN FROM EMBRYONIC STEM CELLS VIA BLASTOCYST COMPLEMENTATION IN MIC. Research Project ...
A blastocyst that had a collapsed blastocoel also showed high levels of TRP53 compared to morphologically normal blastocysts. ... Morphologically normal blastocysts tended to show little nuclear accumulation of stain. However, some cells within these ... Variable expressivity of the tumour suppressor protein TRP53 in cryopreserved human blastocysts. Overview of attention for ... Variable expressivity of the tumour suppressor protein TRP53 in cryopreserved human blastocysts ...
Furthermore, potentially blastocyst-specific miRNAs were identified. In pre-implantation embryos, numerous distinct isomiRs ... We aimed at revealing the sex-specific miRNA expression profile of single porcine blastocysts developing in gilts orally ... affected the miRNA profile in blastocysts despite the distinct differential mRNA expression and DNA methylation found in ... the sex of the embryo nor a maternal E2 exposure affected the miRNA expression profile of developing porcine blastocysts. The ...
Blastocyst transfer and fertility treatment Risks of blastocyst transfer Blastocyst photos at different stages of development ... A recent breakthrough for in vitro fertilization is the use of blastocysts. A blastocyst would be implanted five to six days ... This is then known as the blastocyst. The side of the blastocyst where the inner cellular mass (ICM) forms is referred to as ... into the uterus where the blastocysts are inserted into the womb. Blastocysts also offer an advantage because they can be used ...
Blastocyst Freeze Media, Blastocyst Thaw Media, Embryo Freeze Media, Embryo Thaw Media, Complete Early Cleavage Medium® (ECM®) ... blastocyst stage has become a real option after researchers achieved pregnancy rates that were as good as those for blastocysts ... zygotes through day 3 cleavage and blastocyst stage embryos within the fertility area. ...
HATCHING OF THE BLASTOCYST : HATCHING OF THE BLASTOCYST Formation of blastocyst (D4) Shedding of zona pellucida & hatching of ... Implantation of Blastocyst Implantation of Blastocyst : Implantation of Blastocyst Day - 4: Morula reaches in uterine cavity ... SEQUENCE AFTER FERTILIZATION? 1st cleavage after 30 Hours Morula 3days Blastocyst formation 4th day Blastocyst floats in Uterus ... Blastocyst Implantation Prof. Dr. Saeed Shafi Slide 2: Nadia Suleyman, a 32-year old business executive underwent IVF and ...
A trilaminar embryo (or trilaminary blastoderm, or trilaminar germ disk) is an early stage in the development of triploblastic organisms, which include humans and many other animals. It is an embryo which exists as three different germ layers - the ectoderm, the mesoderm and the endoderm. These layers are arranged on top of each other like a stack of paper, giving rise to the name trilaminar, or "three-layered". These three layers arise early in the third week (after gastrulation) from the epiblast (a portion of the mammalian inner cell mass). Swiss embryology (from UL, UB, and UF) hdisqueembry/triderm01 Embryology at UNSW Notes/week3_4 Overview at edu. ...
blastocyst synonyms, blastocyst pronunciation, blastocyst translation, English dictionary definition of blastocyst. n. The ... blastocyst. Also found in: Thesaurus, Medical, Encyclopedia, Wikipedia.. Related to blastocyst: blastocyst transfer ... However at the ARU, blastocyst culture enables embryos to be "grown" in a lab to the blastocyst stage of development (day five ... The number of embryos or blastocysts transferred in each cycle was captured, but embryo and blastocyst transfers were not ...
expression becomes limited to the ICM in the early blastocyst stage. By the late blastocyst stage, while continuing to express ... are not established until the late blastocyst stage. The expression patterns of NANOG. and GATA6. in the human preimplantation ... and recent discoveries have shed new light on the establishment of the three blastocyst lineages. What is less clear, however, ... results in the formation of a blastocyst with three distinct cell lineages. Only one of these lineages, the epiblast, ...
Blastocyst score affects implantation and pregnancy outcome: towards a single blastocyst transfer. Fertil Steril. 2000;73(6): ... Making the blastocyst: lessons from the mouse Katie Cockburn et al. * Portrait of an oocyte: our obscure origin Roger Gosden et ... while Oct4 expression becomes limited to the ICM in the early blastocyst stage. By the late blastocyst stage, while continuing ... Making the blastocyst: lessons from the mouse. Katie Cockburn and Janet Rossant Department of Molecular Genetics, University of ...
Find blastocyst Stock Images in HD and millions of other royalty-free stock photos, illustrations, and vectors in the ... Blastocyst stock photos. 850 Blastocyst stock photos, vectors, and illustrations are available royalty-free. See blastocyst ... blastocyst. Human blastocyst, with inner cell mass. the mammalian conceptus in the post-morula stage, consisting of the ... blastocyst. Human blastocyst, with inner cell mass. the mammalian conceptus in the post-morula stage, consisting of the ...
Human Cloning Obfuscation 8: A Blastocyst, Not an Embryo By Wesley J. Smith * About Wesley J. Smith ... When it is about 7-10 days along, it is a blastocyst. In human biology, the embryo is called a fetus after three months of ... In this edition, bioethicist Jonathan Moreno claims that a blastocyst isnt an embryo. From his Huffington Post column:. ... The egg "reprograms" the DNA in the donor cell to an embryonic state and divides until it has reached the early, blastocyst ...
Blastocyst Injection. The ability of mouse pre-implantation embryos to incorporate cells from other embryos is fundamental to ... Genetically modified ES cells will be injected into C57Bl/6 or C57Bl/6 albino blastocysts. Injected blasts will then be ... ES cell clone contributing to the germ-line is highly dependent on the quality of the ES cells injected into the blastocyst. It ...
2012) Cell Lineage Allocation Within the Inner Cell Mass of the Mouse Blastocyst. In: Kubiak J. (eds) Mouse Development. ... Artus J, Piliszek A, Hadjantonakis AK (2011) The primitive endoderm lineage of the mouse blastocyst: sequential transcription ... Rossant J, Tam PP (2009) Blastocyst lineage formation, early embryonic asymmetries and axis patterning in the mouse. ... Cell Lineage Allocation Within the Inner Cell Mass of the Mouse Blastocyst. ...
Blastocyst transcription of ,i,OCT4,/i, (pluripotency marker) was not affected by LPA stimulation. In conclusion, LPA is an ... between Days 2 and 8 had no effect on embryo yield and quality and on blastocyst relative mRNA abundance of genes involved in ... Transcription levels of genes coding for LPA synthesis enzymes ((a) ATX; (b) cPLA 2) in late cleavage (Day 5) and blastocyst ( ... Blastocyst transcription of OCT4 (pluripotency marker) was not affected by LPA stimulation. In conclusion, LPA is an early ...
I have just had a grade 5CC blastocyst and an early blastocyst transferred yesterday. My egg quality has been an issue for me, ... I have just had a grade 5CC blastocyst and an early blastocyst transferred yesterday. My egg quality has been an issue for me, ... I feel like my blastocyst is a lost cause, but then I hear stories like yours and it gives me a glimmer of hope. I hope you ...
For one thing, blastocysts have a better chance of implantation, which means the success rate with blastocyst transfers is ... Also, since the doctor is transferring only 1 - 2 blastocysts, you can freeze the extra blastocysts , which means you get a ... till they become blastocysts. A blastocyst is the final stage of the embryos development before it hatches out of its shell ( ... Why you should insist that your doctor do only blastocyst transfers. In the past, most embryo transfers were done on day two or ...
Derivation of embryonic stem-cell lines from human blastocysts.. Cowan CA1, Klimanskaya I, McMahon J, Atienza J, Witmyer J, ...
This mimics findings in OCT4-deficient human blastocysts but is in sharp contrast to Oct4-null mouse blastocysts, where NANOG ... OCT4/POU5F1 is required for NANOG expression in bovine blastocysts. Kilian Simmet, Valeri Zakhartchenko, Julia Philippou- ... OCT4/POU5F1 is required for NANOG expression in bovine blastocysts. Kilian Simmet, Valeri Zakhartchenko, Julia Philippou- ... Control blastocysts showed a typical salt-and-pepper distribution of NANOG- and GATA6-positive cells in the ICM. In contrast, ...
Day 5 blastocyst embryo transfer vs. day 3 cleavage stage transfer is discussed. Advanced Fertility Center of Chicago has done ... Cost for blastocyst transfer. We do not charge extra for blastocyst transfer. See our current fees for IVF ... Day 5 Blastocyst Transfer for IVF. 2 blastocyst embryos that were transferred with a pregnancy resulting. Transferring 2 ... Unresolved issues regarding blastocyst transfer:. *How do we select the most appropriate candidates for blastocyst transfer? ...
Activating genes for reprogramming factors for a short time transforms large numbers of differentiated cells into multipotent forms that could be useful for cell-based therapies.. 0 Comments. ...
The preimplantation mouse blastocyst. A schematic drawing of the differentiated mouse blastocyst. The inner cell mass (ICM) and ... The blastocyst lane contain ≈279 blastocysts for a total of ≈7 μg of protein. ... female mice 48 h after mating and cultured to a blastocyst stage as described (1). The blastocysts were fixed on glass slides ... Blastocysts from each condition were incubated in medium at a final glucose concentration of 5.6 mM with 500 nM insulin (Sigma ...
When the couple produce enough blastocysts of good quality, the selection of the euploid blastocyst for transfer is beneficial ... Study of the Best Blastocyst Post Transfer by aCGH. The safety and scientific validity of this study is the responsibility of ... Study of the Best Blastocyst Post Transfer by aCGH. Brief Summary An Observational, blind and prospective study of ... Other important developments include the culture of pre-embryos to the blastocyst stage. This allows more cells to be obtained ...
... is now offering preimplantation genetic diagnosis testing of blastocyst embryos. The testing identifies genetic defects in ... is now offering preimplantation genetic diagnosis testing of day 5 blastocyst embryos. The testing identifies genetic defects ... Now Offering Trophectoderm Preimplantation Genetic Diagnosis Testing of Blastocyst Embryos. ...
Observation that the blastocysts collapsed after biopsy, yet maintained viability, led to the hypothesis that the collapsed ... Biopsies were obtained without affecting the pregnancy rate of the blastocysts after transfer. Genetic analysis after whole ... Further work showed that vitrification of collapsed blastocysts using an ethylene glycol-containing medium was associated with ... we developed a technique to puncture the capsule and obtain cells from the trophoblast of expanded equine blastocysts, using ...
The cells inside the blastocyst are called the inner cell mass and give rise to the head, body, and other structures vital to ... By 4 to 5 days, a cavity forms within this ball of cells and the embryo is then called a blastocyst. ... The Morula and Blastocyst. Select Subtitle Language. 88 languages available. Afrikaans. Albanian (Tosk) [Shqip]. Arabic [عربي] ...
For example, a blastocyst quality grade of 4AB means that the blastocyst is expanded (grade 4), has many tightly packed cells ... This blastocyst grading system assigns 3 separate quality scores to each blastocyst embryo: ... Blastocyst Hatching out of Zona (shell) = Grade 5. (I) Inner Cell mass = Grade A (Very Good). (T) Trophectoderm = Grade A (Very ... Blastocyst completely hatched = Grade 6. (I) Inner Cell mass = Grade A (Very Good). (T) Trophectoderm = Grade A (Very Good) ...
Study of the Best Blastocyst Post Transfer by aCGH. The recruitment status of this study is unknown. The completion date has ... Clinical pregnancy [ Time Frame: Five weeks after blastocyst transfer ]. Gestational sac with a heartbeat ... Trophectoderm molecular karyotype [ Time Frame: After blastocyst transfer ]. Trophectoderm ploidy Imbalanced chromosome ...
The mammalian blastocyst is a hollow ball of cells containing two cell types, the inner cell mass and the trophectoderm[GO]. ... The mammalian blastocyst is a hollow ball of cells containing two cell types, the inner cell mass and the trophectoderm[GO]. ... Gilbert fig11.32 has blastocyst has giving rise to ICM and trophoblast (which in this source is a synonym for trophectoderm) ... Editors note: Gilbert fig11.32 has blastocyst has giving rise to ICM and trophoblast (which in this source is a synonym for ...
What is blastocyst transfer? Meaning of blastocyst transfer as a legal term. What does blastocyst transfer mean in law? ... Definition of blastocyst transfer in the Legal Dictionary - by Free online English dictionary and encyclopedia. ... Blastocyst Stage Cryopreservation: Blastocyst freezing has three major rationales: (1) the superiority of blastocyst-stage over ... Overall Blastocyst Quality, Trophectoderm Grade, and Inner Cell Mass Grade Predict Pregnancy Outcome in Euploid Blastocyst ...
Home > Groups > Getting Pregnant > Fertility Treatments > Only one 5day blastocyst so far ... As of day 7, we have one more blastocyst, hatching 5BB, for PGS. So two total (4BB and 5BB) sent for PGS testing today with ... Obviously hoping desperately for all three to grow to blastocysts day 6. Then we are send for pgs testing, for hopeful FET next ... 10 embryos on day 3, which has dwindled to one 4BB blastocyst today day 5. Three more compacting as of today. ...
  • The blastocyst consists of cells forming an outer trophectoderm (TE, trophoblast ) layer, an inner cell mass (ICM, embryo blast) and a blastocoel (fluid-filled cavity). (edu.au)
  • A blastocyst is a fertilized egg that has developed for 5-6 days and contains 3 distinct features including a fluid-filled cavity trophectoderm/trophoblast (T) cells, and an inner cell mass (ICM). (fertilitysmarts.com)
  • In this study, the effects of artificial collapseand reduction of the fluid volume in blastocyst cavity before vitrification process on the survival rate and quality of blastocysts were assessed by estimating the expression levels of Oct4 , Nanog , Sox2 , and Klf4 . (reproduction-abstracts.org)
  • The developing embryo is defined as a blastocyst from the appearance of the blastocoele cavity, and now contains two cell populations- the surrounding outer trophoblast cells, and the inner cell mass cells, located at one side of the inner cavity. (lifemapsc.com)
  • Establishment of embryonic stem cells from rat blastocysts. (nih.gov)
  • Generation of Blastocyst-like Structures from Mouse Embryonic and Adult Cell Cultures. (bioportfolio.com)
  • The CDMLe012-A-1 human embryonic stem cell (hESC) line, derived from a day six blastocyst with a normal 46,XX female karyotype spontaneously lost an X-chromosome during cell culture. (bioportfolio.com)
  • The establishment of embryonic stem-cell lines was performed using the hatching blastocysts that were cultured in the presence of IGF-1 or IGFBP-1/IGF-1. (nih.gov)
  • With blastocyst implantation, unlike a day three embryo which must continue to develop following embryo transfer, a transferred blastocyst will implant much more quickly. (londonwomensclinic.com)
  • The blastocyst transfer or " Day5 Transfer " allows fertilized eggs to grow for an additional 2 days past the standard three-day incubation to the point where it is ready to implant. (sdfertility.com)
  • Antiserum prepared in rabbit against 4-day-old mouse cerebellum (anti-NS-4 serum) reacts in the complement-mediated cytotoxicity test with unfertilized and fertilized mouse eggs, cleavage stage embryos, and cells of the trophoblast and inner cell mass of the mouse blastocyst. (researchwithnj.com)
  • A single mouse blastomere from an embryo until the 8-cell stage can generate an entire blastocyst. (bioportfolio.com)
  • The blastocyst stage and the subsequent implantation are critical for a successful pregnancy, yet are challenging to study in vivo. (bioportfolio.com)
  • However, the basic rule of thumb is that the best embryos make it to the blastocyst stage, and therefore has a greater chance statistically of producing an ongoing pregnancy than a lesser quality one. (londonwomensclinic.com)
  • Once the embryo reaches the blastocyst stage, the embryologist can better evaluate the quality of the embryo and to increase the chance of implanting successfully resulting in higher success rates. (sdfertility.com)
  • In humans, blastocyst stage of development occurs during the first and second week following fertilization ( GA week 3 and 4) and is described initially as Carnegie stage 3 . (edu.au)
  • This stage is followed by blastocyst hatching and implantation. (edu.au)
  • This is when the embryo - at blastocyst stage - fixes itself to an inner layer of the uterus called the endometrium. (theduff.co.uk)
  • To avoid this situation the embryos are left to grow for 2or3 days more to reach the 5 or 6 day stage and a blastocyst transfer is performed. (internationalfertilitycentre.com)
  • Most of the fertilized eggs develop into a 3 day old embryo, but roughly half of them reach to the 5th day or the blastocyst stage. (internationalfertilitycentre.com)
  • In order to enumerate the total cell number of cultured embryos, the embryos that progressed to the blastocyst stage for both groups were cultured in Hoechst H33342 medium (5 μg /ml) for three to five minutes, and then mounted onto a slide and examined under a fluorescence microscope. (nih.gov)
  • About 20 percent showed continued development, some all the way to the blastocyst stage, demonstrating the viability of the manipulated embryos. (the-scientist.com)
  • The team found that 47 percent (eight out of 17) of the control group embryos survived through the blastocyst stage, compared to 19 percent of the CRISPR-edited ones. (laboratoryequipment.com)
  • I have shown that in the preimplantation embryo, SNAI1 and SNAI2 have a unique asymmetric distribution pattern from the 2-cell to 8-cell stage, and are then segregated to the TE of the blastocyst. (uwo.ca)
  • The zygote contains both pro-nuclei and is contained within the zona pellucida, until the blastocyst stage. (lifemapsc.com)
  • C. The blastocyst stage. (lifemapsc.com)
  • In the blastocyst stage, the embryo hatches from the zona pellucida layer and is now able to begin implantation into the maternal endometrium. (lifemapsc.com)
  • Four experiments were designed to evaluate the overall and temporal effects of conditioned medium (CM) by bovine cumulus cells on development of nuclear transfer (NT) bovine embryos and to examine the chromosomal composition and allocation of inner cell mass (ICM) and trophectoderm (TE) of the subsequent blastocysts. (usu.edu)
  • These findings suggest that targeting OCT4 in human embryos reduces both viability and quality of blastocysts," the team wrote in the study. (laboratoryequipment.com)
  • This means that all the stuff mentioned above will have happened in the lab rather than in your fallopian tube, and then the blastocyst will be transferred directly into your uterus on day 5. (theduff.co.uk)
  • Implantation represents attaching of the blastocyst to the shell of the uterus, where it then starts do dig in. (healthypair.net)
  • Blastocyst usually nests in the upper half of the uterus on the front or rear shell. (healthypair.net)
  • Inner cells on that thickened spot on the blastocyst start do develop into an embryo, and the outer cells dig into the uterus shell, forming placenta. (healthypair.net)
  • This is a minor bleeding, commonly accompanied by cramps in the lower part of the stomach, which is caused by contraction of the uterus muscles as the blastocyst is finding its way into it. (healthypair.net)
  • Physiological profile of undifferentiated bovine blastocyst-derived trophoblasts [7] "Trophectoderm of blastocysts mediate early events in fetal-maternal communication, enabling implantation and establishment of a functional placenta. (edu.au)
  • Within the blastocyst, the group of cells "decide" to remain as part of the growing embryo or become one of two other structures¾the placenta or yolk sac. (laboratoryequipment.com)
  • Materials & Methods: Mouse blastocysts divided into 5 groups including: A) Vitrified- thawed blastocysts, B) Vitrified- thawed blastocysts after artificial collapse, C) Collapsed blastocysts, D) Immersed blastocysts in vitrification/warming solutions and E) Fresh blastocysts as control group. (reproduction-abstracts.org)
  • It can be concluded that artificial collapse of blastocyst could be a simple and effective way to contribute to the successful blastocyst vitrification. (reproduction-abstracts.org)
  • The preimplantation embryo culminates in the development of a fluid filled structure called the blastocyst. (uwo.ca)
  • While lab-developed embryos may not have enough time to fully demonstrate their overall viability, more advanced blastocysts can. (fertilitysmarts.com)
  • In this study, we developed methods for culturing undifferentiated bovine blastocyst-derived trophoblasts and used both transcriptomics and proteomics in early colonies to categorize and elucidate their functional characteristics. (edu.au)
  • In conclusion, bovine cumulus cell-CM and CR1aa with co-culture supported comparable development and blastocyst ICM:total cell ratio of bovine NT embryos. (usu.edu)
  • But, as the researchers report in Nature , the blastocyst is incapable of fully forming without OCT4, and essentially implodes. (laboratoryequipment.com)
  • A blastocyst that had a collapsed blastocoel also showed high levels of TRP53 compared to morphologically normal blastocysts. (altmetric.com)
  • One of the first key developmental patterning decisions in the morula to blastocyst development is TE or ICM cell fate. (edu.au)
  • A blastocyst differs from an embryo because of its advanced cell development, differentiation, and growth. (fertilitysmarts.com)
  • The 24 hr-conditioned BCM yielded higher blastocyst development than 48 and 72 hr-conditioned BCM. (usu.edu)
  • Morula and blastocyst development were not different among the groups cultured in BCM for 72, 96, and 168 hr, but were significantly higher (P (usu.edu)
  • Zygote vs embryo, blastocyst and foetus: whats the difference? (theduff.co.uk)
  • What's the difference between a zygote, embryo, blastocyst and foetus? (theduff.co.uk)
  • Starting from a single stem cell type, extended pluripotent stem (EPS) cells, we established a 3D differentiation system that enabled the generation of blastocyst-like structures (EPS-blastoids) through lineage segregation and self-organization. (bioportfolio.com)
  • Building Blastocysts from Stem Cells. (bioportfolio.com)
  • 2019) describe a novel way to generate blastocyst-like structures only from pluripotent stem cells. (bioportfolio.com)
  • Shell of the blastocyst is made by one layer of cells in all but one area where three to four layers of cells are found. (healthypair.net)
  • Those couples who have undergone unsuccessful IVF cycles or IVF/ICSI in-spite of having good quality eggs are the best candidates for a blastocyst culture. (internationalfertilitycentre.com)
  • Recent reports show that more fertility clinics in the country implement the procedure of blastocyst transfers to decrease the risk of multiple births from IVF . (sdfertility.com)
  • Learn more about blastocyst transfers . (sdfertility.com)
  • EPS-blastoids resembled blastocysts in morphology and cell-lineage allocation a. (bioportfolio.com)
  • We have published hundreds of Generation Functional Lungs Conditional Blastocyst Complementation Using Pluripotent news stories on BioPortfolio along with dozens of Generation Functional Lungs Conditional Blastocyst Complementation Using Pluripotent Clinical Trials and PubMed Articles about Generation Functional Lungs Conditional Blastocyst Complementation Using Pluripotent for you to read. (bioportfolio.com)
  • In addition to the medical data, news and clinical trials, BioPortfolio also has a large collection of Generation Functional Lungs Conditional Blastocyst Complementation Using Pluripotent Companies in our database. (bioportfolio.com)
  • IGF-1/IGFBP-1 increases blastocyst formation and total blastocyst cell number in mouse embryo culture and facilitates the establishment of a stem-cell line. (nih.gov)
  • Mouse blastocyst immunosurgery with commercial antiserum to mouse eryt" by Yolanda P. Cruz, Robin S. Treichel et al. (oberlin.edu)
  • Morphologically normal blastocysts tended to show little nuclear accumulation of stain. (altmetric.com)