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  • embryos
  • We applied this system to stably modify chicken primordial germ cell (PGC) lines in vitro and induce a reporter gene at specific developmental stages after injection of the transposon-modified germ cells into chicken embryos. (jove.com)
  • Full understanding of post-transcriptional regulation of gene expression in early embryos is crucial to understand the reprogramming of the embryonic genome, it might help driving reprogramming of stem cells in vitro and will likely improve in vitro culturing of mammalian embryos for assisted reproduction. (mdpi.com)
  • Using wildtype or engineered stem cell lines, researchers may use this technique to uncover the various mechanisms or treatments that may affect early brain infection and resulting microcephaly in Zika virus-infected embryos. (jove.com)
  • There are also methods for PCR-based cloning, differentiating embryonic stem cells into embryonic bodies, live imaging of mammalian embryos using multiphoton microscopy, and the cloning of pig, cattle, rabbit, and mouse oocytes. (springer.com)
  • Knowledge about how these lineages develop during the preimplantation period has major clinical implications for increasing the success of assisted reproductive strategies (ARTs) such as in vitro fertilization (IVF) and preimplantation genetic diagnosis (PGD), preventing the high rate of early pregnancy loss in humans, and improving the derivation of stem cell lines from human embryos. (jci.org)
  • Effect of glycosaminoglycans on in vitro fertilizing ability and in vitro developmental potential of bovine embryos. (thefreelibrary.com)
  • S.v. Effect of glycosaminoglycans on in vitro fertilizing ability and in vitro developmental potential of bovine embryos. (thefreelibrary.com)
  • Stem cell biology has been the subject of much recent discussion, but only the totipotent fertilized oocyte and blastomere cells of embryos at the 2- to 8-cell stage are capable of generating a fully viable organism. (ahajournals.org)
  • However, when a somatic cell nucleus is transplanted into an intact oocyte containing its own chromosomes, the resulting polyploid embryos are able to develop to blastocysts and support ESC derivation. (pubmedcentralcanada.ca)
  • Based on the results obtained for the studies of in vitro development of cloned embryos, epigenetic modifications have been widely used for cloning farm animals. (reproduction.jp)
  • Suberoylanilide hydroxamic acid during in vitro culture improves development of dog-pig interspecies cloned embryos but not dog cloned embryos. (reproduction.jp)
  • Porcine oocytes supported reprogramming of nuclei from dog fibroblasts up to early developmental stage of iSCNT embryos, and treating the embryos with suberoylanilide hydroxamic acid (SAHA) increased their developmental competence. (reproduction.jp)
  • Because the ectopic transplantation of normal peri-implantation embryos can give rise to pluripotent cell lines, the direct derivation of pluripotent cell lines in vitro was attempted without the teratocarcinoma step. (biologists.org)
  • It is well known that in vitro fertilized (IVF) embryos are less competent compared to embryos produced in vivo [ 21 , 22 ]. (ejast.org)
  • In spite of the efforts made to improve these suboptimal conditions, in vitro produced (IVP) embryos are still significantly affected, whether measured in terms of morphology, apoptotic index, blastocyst rate, cryotolerance, gene expression or postnatal growth [ 22 ]. (ejast.org)
  • differentiation
  • During early differentiation the number of cells expressing Oct4 (potential germ-like cells) reaches ~5%, however currently the formation of OLCs remains relatively inefficient. (jove.com)
  • To investigate the differentiation ability of human adipose-derived stem cells (ASCs) towards urothelium-like cells in vitro and the dynamic changes of related cytokines and cytokine receptors in the culture medium. (jove.com)
  • Despite the identification and isolation of glial progenitors from adult human sub-cortical white matter and development of various culture conditions to direct differentiation of fetal neural progenitors into myelin producing oligodendrocytes, acquiring sufficient human oligodendrocytes for in vitro experimentation remains difficult. (jove.com)
  • New naturally occurring mutations in genes for the oocyte-derived growth factors growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15) are described. (las.ac.cn)
  • The authors found that STAT3 kockout (KO) did not affect the differentiation of human ESCs into human intestinal organoids (hIOs) but rather affected the in vitro maturation of hIOs. (escellnews.com)
  • Oct4 is required for ES cell self-renewal, and is highly expressed in ES cells and quickly down-regulated during differentiation 6,7 . (jove.com)
  • Considerable work remains to further define the requirements for in vitro differentiation of ES cells into mature gametes so that these techniques can be clinically applied in regenerative reproductive medicine protocols. (biomedcentral.com)
  • Protocols for the in vitro differentiation of ES cells into cardiomyocytes representing all specialized cell types of the heart, such as atrial-like, ventricular-like, sinus nodal-like, and Purkinje-like cells, have been established. (ahajournals.org)
  • This review summarizes the current state of ES cell-derived cardiogenesis and provides an overview of how genomic strategies coupled with this in vitro differentiation system can be applied to cardiac research. (ahajournals.org)
  • immature
  • Recombinant tUCHL1 absorbed p34(cdc2), a component of MPF, in obviously larger quantities from mature oocytes than from immature oocytes, and p13(suc1) was isolated from tUCHL1 with a dependence on the ATP regeneration system, suggesting that still other functions may be involved in their association that require phosphorylation. (jove.com)
  • The ATSCs and several immature cells were routinely expressing Oct4 protein before and after differentiating into specific lineages. (jove.com)
  • STAT3 KO hIOs displayed immature morphologies with decreased size and reduced budding in hIOs even after in vitro maturation. (escellnews.com)
  • Recent experimental data showed that ovarian surface epithelium and cortical tissue from both mouse and human were proved to contain very low proportion of cells able to propagate themselves, but also to generate immature oocytes in vitro or in vivo, when transplanted into immunodeficient mice ovaries. (biomedcentral.com)
  • genes
  • Moreover, LPA reduced the occurrence of apoptosis in COCs and promoted an antiapoptotic balance in the transcription of genes involved in apoptosis ( BAX and BCL2 ) either in oocytes or blastocysts. (biomedcentral.com)
  • To clarify the role of the POU domain transcription factor Oct4 in Adipose Tissue Stromal Cells (ATSCs), we investigated the regulation of Oct4 expression and other embryonic genes in fully differentiated cells, in addition to identifying expression at the gene and protein levels. (jove.com)
  • They produced a mutant semi-cloned mice pool by oocyte injection of AG-haESCs carrying constitutively expressed Cas9 and an single guide RNA library targeting 72 preselected genes in one step and screened for bone-development-related genes through skeletal analysis at birth. (escellnews.com)
  • Oocyte-specific gene Oog1 suppresses the expression of spermatogenesis-specific genes in oocytes, pp. 297-301). (reproduction.jp)
  • These findings suggested that OOG1 suppresses the expression of spermatogenesis-specific genes in the oocytes and plays important roles during oogenesis. (reproduction.jp)
  • Cells
  • During FSH-induced maturation, rapid cAMP surges were observed in both cumulus cells and oocyte. (jove.com)
  • Most GJC between cumulus cells and oocyte ceased immediately after FSH stimulation and recommenced after the cAMP surge. (jove.com)
  • These cells could then be differentiated into the affected cell type of the patient and studied in vitro, or used to replace the damaged or missing cells. (jove.com)
  • The availability of various GFP-tagged proteins means that many cellular processes can be monitored in living cells over the course of minutes or hours, and using this technique, processes such as RNP transport, epithelial morphogenesis, and tissue remodeling have been described in great detail in Drosophila oocytes 1,2 . (jove.com)
  • The formation of oocyte-like cells (OLCs) from somatic stem cells, isolated from newborn mouse skin, has been demonstrated and can be visualized in this video protocol. (jove.com)
  • Culturing the normal cells as a primary culture in vitro will enable scientists to model specific changes that might lead to ovarian cancer in the distinct epithelium, thereby definitively determining the cell type of origin. (jove.com)
  • Maintaining normal cells in alginate hydrogels promotes short term in vitro culture of cells in their three-dimensional context and permits introduction of plasmid DNA, siRNA, and small molecules. (jove.com)
  • Here we describe the use of one such method to image bacterial cells harboring the fluorescently-labelled cytokinetic FtsZ protein to show how cells are analyzed and the type of unique information that this technique can provide. (jove.com)
  • Here, the isolated cells are then analyzed using a matrigel invasion assay to assess trophoblast invasiveness in vitro 5-6 . (jove.com)
  • Various cell types could be analyzed using this technique, including lymphocytes/leukocytes, stem cells, and tumor cells. (jove.com)
  • We used piggyBac to gene-modify cytotoxic T lymphocytes to carry a chimeric antigen receptor directed against the tumor antigen HER2, and found that gene-modified T cells mediated targeted killing of HER2-positive tumor cells in vitro and in vivo in an orthotopic mouse model 14 . (jove.com)
  • Oocytes and spermatozoa are terminally differentiated haploid cells which can no longer divide. (mdpi.com)
  • In the cow, lysophosphatidic acid (LPA) acts as an auto-/paracrine factor, through its receptors LPAR1-4, on oocytes and cumulus cells during in vitro maturation (IVM). (biomedcentral.com)
  • Oocytes, cumulus cells and blastocysts were used for gene expression analysis. (biomedcentral.com)
  • EGFP positive cells, has high expression in OCT4 , but a different level of NANOG expression. (jove.com)
  • Human embryonic stem cells (hESCs) are pluripotent and an invaluable cellular sources for in vitro disease modeling and regenerative medicine 1 . (jove.com)
  • In Germ Cell Protocols, expert laboratory investigators describe in step-by-step detail a series of powerful techniques for the molecular and genetic analysis of germ cells in a variety of different reproductive systems. (springer.com)
  • Following three-four days of culture, cells were stained for the pluripotency marker OCT4 and the G2 cell-cycle marker Cyclin B1, and nuclei and cytoplasm were co-stained with Hoechst and Cell Mask Blue, respectively. (escellnews.com)
  • All ICM cells contain Oct4, a key transcription factor that is first expressed at the morula stage. (jove.com)
  • The Oct4GiP cells express the green fluorescent protein (GFP) under the control of the Oct4 gene promoter region 4,5 . (jove.com)
  • The ability of hAFCs to differentiate into germ cell and oocyte-like cells has been previously documented. (biomedcentral.com)
  • Moreover, culture of EBs in medium containing human follicular fluid (HFF) or a germ cell maturation factor cocktail (FAC), expressed germ cells markers such as BLIMP1 , STELLA , DAZL , VASA , STRA8 , SCP3 , SCP1 , and GDF9 . (biomedcentral.com)
  • Nonetheless, while Dazl expression was required for deriving germ cells from murine ES cells in vitro, this only supported progression through the early stages of meiosis. (biomedcentral.com)
  • By cultivation in vitro as 3D aggregates called embryoid bodies, ES cells can differentiate into derivatives of all 3 primary germ layers, including cardiomyocytes. (ahajournals.org)
  • Methods of producing human stem cells are disclosed for parthenogenetically activating human oocytes by manipulation of O 2 tension, including manipulation of Ca 2 + under high O 2 tension and contacting oocytes with serine threonine kinase inhibitors under low O 2 tension, isolating inner cell masses (ICMs) from the activated oocytes, and culturing the cells of the isolated ICMs under high O 2 tension. (patentsencyclopedia.com)
  • Moreover, methods are described for the production of stems cells from activated oocytes in the absence of non-human animal products, including the use of human feeder cells/products for culturing ICM/stem cells. (patentsencyclopedia.com)
  • 90. A method of treating a subject in need thereof, comprising administering a cellular composition comprising differentiated cells, wherein the differentiated cells are derived from a stem cell obtained from a parthenogenetically activated oocyte from a human donor. (patentsencyclopedia.com)
  • Confocal laser scanning microscopy indicated a higher number of transzonal processes (TZPs) reaching the oocyte from cumulus cells in 2% PVP medium than in 0% PVP medium. (reproduction.jp)
  • Whether cumulus cells are required for the successful vitrification of mature oocytes in cattle is controversial. (reproduction.jp)
  • re-evaluated the effects of the presence of multi-layered cumulus cells (MCCs) on the vitrification of mature bovine oocytes (Ishii et al . (reproduction.jp)
  • Eventually, conditions were developed that allowed the culture of EC cells in vitro, establishing them as an in vitro model of mammalian development ( Kahan and Ephrussi, 1970 ). (biologists.org)
  • Oocytes of women affected by metabolic disorders, such as diabetes or obesity and oocyte aging, can be improved by transfer of mitochondria from cells with mitochondrial integrity into mitochondria-impaired oocytes. (biomedcentral.com)
  • Deepa Bhartiya and colleagues [ 9 ] reviewed in vitro approaches for the development of oocytes and sperm from embryonic stem cells (ESC), induced pluripotent stem cells (iPS cells), ovarian stem cells (OSCs), pluripotent stem cells (PSCs), spermatogonial stem cells (SSCs), and very small embryonic-like stem cells (VSELs). (biomedcentral.com)
  • ovarian
  • The ovarian follicle is the functional unit of the ovary that secretes sex hormones and supports oocyte maturation. (jove.com)
  • Our in vitro culture system employs hydrogels in order to mimic the native ovarian environment by maintaining the 3D follicular architecture, cell-cell interactions and paracrine signaling that direct follicle development 5 . (jove.com)
  • Herein, we describe an encapsulation method and an in vitro culture system for ovarian follicles within a FA-IPN. (jove.com)
  • Although anandamide (AEA) had been measured in human follicular fluid and is suggested to play a role in ovarian follicle and oocyte maturity, its exact source and role in the human ovary remains unclear. (jove.com)
  • For many years, ovarian biology has been based on the dogma that oocytes reserve in female mammals included a finite number, established before or at birth and it is determined by the number and quality of primordial follicles developed during the neonatal period. (biomedcentral.com)
  • stem cell
  • Co-expression of cyp26a1 and the pluripotent germ cell stem cell marker pou5f1(oct4) in meiotically arrested oocytes was consistent with roles in mouse to promote germ cell survival and to prevent apoptosis, mechanisms that are central for tipping the sexual fate of gonads towards the female pathway in zebrafish. (jove.com)
  • Therefore stem cell therapy may potentially provide an avenue to treat women with POF, reproductive aging and/or poor oocyte quality. (biomedcentral.com)
  • 60. A differentiated cell derived from a stem cell obtained from a parthenogenetically activated oocyte from a human donor. (patentsencyclopedia.com)
  • The ability to control the transition from an undifferentiated stem cell to a specific cell fate is one of the key techniques that are required for the application of interventional technologies to regenerative medicine and the treatment of tumors and metastases and of neurodegenerative diseases. (springer.com)
  • germ
  • Currently the term "germ plasm" refers to a large complex of proteins and RNA that visually polarizes the oocyte. (stembook.org)
  • meiotic arrest
  • Many studies have shown that cyclic adenosine-5-monophosphate (cAMP)-dependent protein kinase A (PKA) and G-protein-coupled receptor 3 (GPR3) are crucial for controlling meiotic arrest in oocytes. (jove.com)
  • Consistent with this hypothesis, females up-regulated cyp26a1 in oocytes that entered prophase-I meiotic arrest, and down-regulated cyp26a1 in oocytes resuming meiosis. (jove.com)
  • follicles
  • Isolated follicles can be used for a variety of experimental techniques, including in vitro development assays, live imaging, mRNA expression analysis and western blot analysis of proteins. (jove.com)
  • Restored ovaries displayed many follicle-enclosed oocytes at all stages of development, but no oocytes or follicles were observed in sterilized mice whose ovaries had been injected with medium only (control). (biomedcentral.com)
  • mice
  • Adenovirus-mediated Gene Delivery and in Vitro Microinsemination Produce Offspring from Infertile Male Mice Proceedings of the National Academy of Sciences of the United States of America. (jove.com)
  • generated Oog1 RNA interference (RNAi)-transgenic mice to investigate the effects of Oog1 knockdown on gene expression in the oocytes (Honda et al . (reproduction.jp)
  • embryonic
  • This oocyte-to-embryo transition needs an efficient and tightly regulated translation of the maternally-inherited mRNAs, which likely contributes to embryonic genome activation. (mdpi.com)
  • In vitro, the epiblast precursor is most closely represented by the well-characterized embryonic stem (ES) cell lines that maintain the expression of Oct4, but analogous ExEn precursor cell lines are not known and it is unclear if they would express Oct4. (jove.com)
  • Cytoplasmic factors present in mature, metaphase II (MII)-arrested oocytes have a unique ability to reset the identity of transplanted somatic cell nuclei to the embryonic state. (pubmedcentralcanada.ca)
  • competent
  • The production of competent oocytes during IVM is important for cattle reproduction concerning the ability to increase production of valuable, healthy offspring, application of nuclear transfer and transgenic technologies for agricultural usage. (biomedcentral.com)
  • induce
  • In the presence of the GJC inhibitor carbenoxolone (CBX), FSH failed to induce the meiotic resumption and the changes in PKAI, GPR3 and cAMP surge in oocyte were no longer detected. (jove.com)
  • OLCs
  • The resulting OLCs express various markers consistent with oocytes such as Oct4 , Vasa , Bmp15 , and Scp3 . (jove.com)
  • SCNT
  • Hence, a technique called Somatic Cell Nuclear Transfer (SCNT) was developed. (blogspot.com)
  • However, despite numerous applications of SCNT for animal cloning, the nature of reprogramming oocyte factors and their mechanism of action remain largely unknown. (pubmedcentralcanada.ca)
  • Though the underlying cause of early developmental arrest remains unclear, most of these studies involving human oocytes applied SCNT protocols developed for nonprimate species. (pubmedcentralcanada.ca)
  • Advanced reproductive technologies, such as somatic cell nuclear transfer (SCNT), challenge us with a new type of embryo, with special nutritional requirements and altered physiology under in vitro conditions. (ejast.org)
  • In general terms, SCNT involves the removal of the nucleus and polar body from an oocyte (cytoplast) that then receives a nucleus from an external somatic cell (donor nucleus). (ejast.org)
  • assays
  • 1988). Moreover, several DNA glycosylases, including TDG and MBD4, have been implicated in DNA demethylation, although none of them has shown convincing activity on 5mC in in vitro enzymatic assays (Kangaspeska et al. (arraystar.com)
  • processes
  • Further investigation of these processes revealed that methylglyoxal may affect the oocyte quality by resulting in excessive reactive oxygen species production, aberrant mitochondrial distribution and high level lipid peroxidation, and resveratrol can block these cytotoxic changes. (jove.com)
  • this allows one to combine genetic or pharmacologic perturbations with in vitro development to define the effects of such manipulations on the processes occurring during specific periods of development. (jove.com)
  • unclear
  • However, it is unclear how gonadotropins modulate these factors to regulate oocyte maturation, especially by gap junctional communication (GJC). (jove.com)
  • Xenopus
  • Additionally, many of the techniques to overexpress or inhibit gene function in organisms such as Xenopus and zebrafish fail to sufficiently target critical maternal signaling pathways, such as Wnt signaling. (jove.com)
  • In Xenopus , manipulating gene function in cultured oocytes and subsequently fertilizing them can ameliorate these problems to some extent. (jove.com)
  • protein
  • Then, we exclude protein availability and ubiquitin carboxyl-terminal hydrolase enzyme activity as factors in the response of oocytes to winter hibernation. (jove.com)
  • In the context of MPF (maturation promoting factor) controlling oocyte maturation and to further understand the role of UCHL1 in oocyte maturation, we performed adsorption and co-immunoprecipitation experiments using toad oocyte protein extracts and determined that tUCHL1 is associated with MPF in toad oocytes. (jove.com)
  • In oocytes from hibernation-interrupted toads, the p34(cdc2) protein level was significantly lower than in oocytes from toads in artificial hibernation, providing an explanation for the different quantities isolated by recombinant tUCHL1 pull-down and, more importantly, identifying a mechanism involved in the toad oocytes dependence on a low environmental temperature during winter hibernation. (jove.com)
  • Oog1 , an oocyte-specific gene, encodes the protein belonging to the leucine-rich repeat (LRR) superfamily. (reproduction.jp)
  • tissue
  • The alginate hydrogel floats in standard cell culture medium and supports the architecture of the tissue growth in vitro . (jove.com)
  • In vitro expansion of neural progenitors from fetal CNS tissue has been well characterized. (jove.com)
  • chromosomes
  • The spermatozoon mostly contributes the other half of the chromosomes and it is specialized to reach and to penetrate the oocyte. (mdpi.com)