0060] The term "cell proliferation control" means a procedure in which cell proliferative capacity is stopped at a desired time, the cell concentration is maintained without significant damage affecting the viability of cells to the cells, and subsequently cell proliferation is restarted at a desired time. In other words, in a method for controlling cell proliferation of the present invention, the proliferation of primate pluripotent stem cells can be inhibited with a maintenance medium for pluripotent stem cells of the present invention, and after a desired period elapsed, the proliferation of primate pluripotent stem cells can be promoted with a culture medium containing glucose. In a method for controlling cell proliferation of the present invention, the growth rate and the morphology of pluripotent stem cells following the procedure in which the proliferation is inhibited with a maintenance medium for pluripotent stem cells of the present invention, and after an inhibition period passed, the ...
Induced pluripotent stem cell technology makes in vitro reprogramming of somatic cells from individuals with various genetic backgrounds possible. By applying this technology, it is possible to produce pluripotent stem cells from biopsy samples of arbitrarily selected individuals with various genetic backgrounds and to subsequently maintain, expand, and stock these cells. From these induced pluripotent stem cells, target cells and tissues can be generated after certain differentiation processes. These target cells/tissues are expected to be useful in regenerative medicine, disease modeling, drug screening, toxicology testing, and proof-of-concept studies in drug development. Therefore, the number of publications concerning induced pluripotent stem cells has recently been increasing rapidly, demonstrating that this technology has begun to infiltrate many aspects of stem cell biology and medical applications. In this review, we discuss the perspectives of induced pluripotent stem cell technology ...
SAN DIEGO, December 15, 2017 - ViaCyte, Inc., a privately-held regenerative medicine company, today announced that the California Institute for Regenerative Medicine (CIRM) approved a grant of $1.4 million to support the initial development of immune-evasive pluripotent stem cell lines. The focus of the project will be to genetically engineer the Companys CyT49 pluripotent stem cell line. ViaCytes proprietary CyT49 cell line is well characterized and has been used to manufacture cell replacement product candidates that have been reviewed and allowed for use in clinical trials by multiple regulatory authorities.. One of the main challenges in developing an off-the-shelf cell therapy is the potential for immune rejection of implanted cells. Genetic engineering of a pluripotent stem cell line may make it possible to generate cell therapies from a single source that will not be rejected by the immune system. For diabetes, pluripotent stem cells have the potential of providing an unlimited supply ...
Since it was first demonstrated that induced pluripotent stem cells (iPS cells) could be derived from mature cells, significant progress has been made in the field of acquisition, characteristics, identification and application of iPS cells. Until now, diverse means have been proven to generate iPS cells successfully in many biological species and more cell types. Meanwhile, researchers continue to target the efficiency of induction. To identify the characteristics of induced pluripotent stem cells and attest to their pluripotency, one must verify the expression of new derived stem cell genes and proteins, doubling times, methylation patterns, teratoma formation, embryoid body formation, viable chimera formation and capacity to differentiate into all cell types. In other words, induced pluripotent stem cells are theoretically similar, or even same, to natural pluripotent stem cells, for instance embryonic stem (ES) cells. Furthermore, iPS cells have the potential to take the place of ES cells ...
Pluripotent stem cells such as induced pluripotent stem cells (iPSCs) and embryonic stem cells (ESCs) form teratomas when transplanted into immunodeficient mice. As teratomas contain all three germ layers (endoderm, mesoderm, ectoderm), teratoma formation assay is widely used as an index of pluripotency (Evans and Kaufman, 1981; Hentze et al., 2009; Gropp et al., 2012). On the other hand, teratoma-forming tumorigenicity also represents a major risk factor impeding potential clinical applications of pluripotent stem cells (Miura et al., 2009; Okano et al., 2013). Recently, we reported that iPSCs derived from naked mole-rat lack teratoma-forming tumorigenicity when engrafted into the testes of non-obese diabetic/severe combined immunodeficient (NOD/SCID) mice due to an ES cell-expressed Ras (ERAS) and Alternative reading frame (ARF)-dependent tumor-suppression mechanism specific to this species (Miyawaki et al., 2016). Here, we describe a method for transplanting pluripotent stem cells into the testes of
Field potential duration (FPD) in human-induced pluripotent stem cell-derived cardiomyocytes (hiPS-CMs), which can express QT interval in an electrocardiogram, is reported to be a useful tool to predict K{sup +} channel and Ca{sup 2+} channel blocker effects on QT interval. However, there is no report showing that this technique can be used to predict multichannel blocker potential for QT prolongation. The aim of this study is to show that FPD from MEA (Multielectrode array) of hiPS-CMs can detect QT prolongation induced by multichannel blockers. hiPS-CMs were seeded onto MEA and FPD was measured for 2 min every 10 min for 30 min after drug exposure for the vehicle and each drug concentration. I{sub Kr} and I{sub Ks} blockers concentration-dependently prolonged corrected FPD (FPDc), whereas Ca{sup 2+} channel blockers concentration-dependently shortened FPDc. Also, the multichannel blockers Amiodarone, Paroxetine, Terfenadine and Citalopram prolonged FPDc in a concentration dependent manner. ...
Epidermal grafting using cells derived from pluripotent stem cells will change the face of this side of regenerative cutaneous medicine. To date, the safety of the graft would be the major unmet deal in order to implement long-term skin grafting. In this context, experiments on large animals appear unavoidable to assess this question and possible rejection. Cellular tools for large animal models should be constructed. In this study, we generated monkey pluripotent stem cell-derived keratinocytes and evaluated their capacities to reconstruct an epidermis, in vitro as well as in vivo. Monkey pluripotent stem cells were differentiated efficiently into keratinocytes able to reconstruct fully epidermis presenting a low level of major histocompatibility complex class-I antigens, opening the way for autologous or allogeneic epidermal long-term grafting. Functional keratinocytes generated from nonhuman primate embryonic stem cells and induced pluripotent stem cells reproduce an in-vitro and in-vivo stratified
TY - JOUR. T1 - Generation and characterization of human induced pluripotent stem cell (hiPSC) lines from an Alzheimers disease (ASUi003-A) and non-demented control (ASUi004-A) patient homozygous for the Apolipoprotein e4 (APOE4) risk variant. AU - Brookhouser, Nicholas. AU - Zhang, Ping. AU - Caselli, Richard John. AU - Kim, Jean J.. AU - Brafman, David A.. PY - 2017. Y1 - 2017. N2 - Although the majority of late-onset Alzheimers disease (AD) patients are labeled sporadic, multiple genetic risk variants have been identified, the most powerful and prevalent of which is the e4 variant of the Apolipoprotein E (APOE) gene. Here, we generated human induced pluripotent stem cell (hiPSC) lines from the peripheral blood mononuclear cells (PBMCs) of a clinically diagnosed AD patient [ASUi003-A] and a non-demented control (NDC) patient [ASUi004-A] homozygous for the APOE4 risk allele. These hiPSCs maintained their original genotype, expressed pluripotency markers, exhibited a normal karyotype, and ...
Title:The Role of MicroRNAs in the Pancreatic Differentiation of Pluripotent Stem Cells. VOLUME: 3 ISSUE: 1. Author(s):Natalie Francis, Melanie Moore, Guy A. Rutter and Chris Burns. Affiliation:Endocrinology Section, Biotherapeutics Department, National Institute of Biological Standards and Control, Blanche Lane, South Mimms, Hertfordshire, EN6 3QG, UK.. Keywords:Differentiation, embryonic stem cells, endoderm, induced pluripotent stem cells, insulin, microRNA, pancreas, type 1 diabetes, type 2 diabetes.. Abstract:The generation of β-cells in vitro is an attractive option for cell therapy treatments for type 1 diabetes and also for the development of more accurate disease models. A number of studies have demonstrated that insulin-expressing cells can be generated by the in vitro differentiation of human pluripotent stem cells. However, to date, these differentiation protocols are often inefficient, time-consuming and highly variable. In many cases, this is a result of an incomplete ...
In 2007, two independent research groups published manuscripts that described successful genetic reprogramming of human adult somatic cells into pluripotent human stem cells.34,35 Although some technical limitations remain, this strategy suggests a promising new avenue for generating pluripotent cell lines that can inform drug development, models of disease, and ultimately, transplantation medicine. These experiments, which are discussed below, were breakthroughs because they used adult somatic cells to create pluripotent stem cells that featured hallmarks of ES cells.. In 2006, Shinya Yamanaka and colleagues at Kyoto University reported that they could use a retroviral expression vector to introduce four important stem cell factors into adult mouse cells and reprogram them to behave like ES cells (see Figure 8.3k).37 They called the reprogrammed cells "iPSCs," for induced pluripotent stem cells. However, iPSCs produced using the original technique failed to produce sperm and egg cells when ...
BACKGROUND: Human-induced pluripotent stem cells (hiPSCs) are a potentially invaluable resource for regenerative medicine, including the in vitro manufacture of blood products. HiPSC-derived red blood cells are an attractive therapeutic option in hematology, yet exhibit unexplained proliferation and enucleation defects that presently preclude such applications. We hypothesised that substantial differential regulation of gene expression during erythroid development accounts for these important differences between hiPSC-derived cells and those from adult or cord-blood progenitors. We thus cultured erythroblasts from each source for transcriptomic analysis to investigate differential gene expression underlying these functional defects. RESULTS: Our high resolution transcriptional view of definitive erythropoiesis captures the regulation of genes relevant to cell-cycle control and confers statistical power to deploy novel bioinformatics methods. Whilst the dynamics of erythroid program elaboration from
Microphysiological systems (MPS) may be able to provide the pharmaceutical industry models that can reflect human physiological responses to improve drug discovery and translational outcomes. With lack of efficacy being the primary cause for drug attrition, developing MPS disease models would help researchers identify novel targets, study mechanisms in more physiologically-relevant depth, screen for novel biomarkers and test/optimize various therapeutics (small molecules, nanoparticles and biologics). Furthermore, with advances in inducible pluripotent stem cell technology (iPSC), pharmaceutical companies can access cells from patients to help recreate specific disease phenotypes in MPS platforms. Combining iPSC and MPS technologies will contribute to our understanding of the complexities of neurodegenerative diseases and of the blood brain barrier (BBB) leading to development of enhanced therapeutics. © 2018. ...
Human-induced pluripotent stem cells (hiPSCs) show a great promise as a renewable source of cells with broad biomedical applications. Since insulin has been used in the maintenance of hiPSCs, in this study we explored the role of insulin in culture of these cells. We report conditions for insulin starvation and stimulation of hiPSCs. Crystal violet staining was used to study the adhesion and proliferation of hiPSCs. Apoptosis and cell cycle assays were performed through flow cytometry. Protein arrays were used to confirm phosphorylation targets, and mRNA sequencing was used to evaluate the effect of transcriptome. Insulin improved the seeding and proliferation of hiPSCs. We also observed an altered cell cycle profile and increase in apoptosis in hiPSCs in the absence of insulin. Furthermore, we confirmed phosphorylation of key components of insulin signaling pathway in the presence of insulin and demonstrated the significant effect of insulin on regulation of the mRNA transcriptome of hiPSCs. Insulin is
This medium-ACF is a serum-free medium designed for optimal growth of human embryonic stem cells and induced pluripotent stem cells under feeder-free conditions. It is a sterile, liquid medium containing essential and non-essential amino acids, vitamins, organic and inorganic compounds, hormones, growth factors and trace minerals. The medium is bicarbonate buffered and has a pH of 7.4 when equilibrated in an incubator with an atmosphere of 5% CO2/ 95% air. This medium-ACF is formulated (quantitatively and qualitatively) to provide a defined and optimally balanced nutritional environment that selectively promotes growth of normal human embryonic stem cells and induced pluripotent stem cells in vitro. This medium should be used in conjunction with BD Matrigel™ for feeder-free culture condition ...
Recent studies have revealed negligible immunogenicity of induced pluripotent stem (iPS) cells in syngeneic mice and in autologous monkeys. Therefore, human iPS cells would not elicit immune responses in the autologous setting. However, given that human leukocyte antigen (HLA)-matched allogeneic iPS cells would likely be used for medical applications, a more faithful model system is needed to reflect HLA-matched allogeneic settings. Here we examined whether iPS cells induce immune responses in the swine leukocyte antigen (SLA)-matched setting. iPS cells were generated from the SLA-defined C1 strain of Clawn miniature swine, which were confirmed to develop teratomas in mice, and transplanted into the testes (n = 4) and ovary (n = 1) of C1 pigs. No teratomas were found in pigs on 47 to 125 days after transplantation. A Mixed lymphocyte reaction revealed that T-cell responses to the transplanted MHC-matched (C1) iPS cells were significantly lower compared to allogeneic cells. The humoral immune responses
TY - JOUR. T1 - SALL3 expression balance underlies lineage biases in human induced pluripotent stem cell differentiation. AU - Kuroda, Takuya. AU - Yasuda, Satoshi. AU - Tachi, Shiori. AU - Matsuyama, Satoko. AU - Kusakawa, Shinji. AU - Tano, Keiko. AU - Miura, Takumi. AU - Matsuyama, Akifumi. AU - Sato, Yoji. PY - 2019/12/1. Y1 - 2019/12/1. N2 - Clinical applications of human induced pluripotent stem cells (hiPSCs) are expected, but hiPSC lines vary in their differentiation propensity. For efficient selection of hiPSC lines suitable for differentiation into desired cell lineages, here we identify SALL3 as a marker to predict differentiation propensity. SALL3 expression in hiPSCs correlates positively with ectoderm differentiation capacity and negatively with mesoderm/endoderm differentiation capacity. Without affecting self-renewal of hiPSCs, SALL3 knockdown inhibits ectoderm differentiation and conversely enhances mesodermal/endodermal differentiation. Similarly, loss- and gain-of-function ...
Methods and Results-Transgenic hiPSC lines stably expressing a fluorescent reporter and NIS (NISpos-hiPSCs) were established. Iodide uptake, efflux, and viability of NISpos-hiPSCs were assessed in vitro. Ten (±2) days after induction of myocardial infarction by transient occlusion of the left anterior descending artery, catheter-based intramyocardial injection of NISpos-hiPSCs guided by 3-dimensional NOGA mapping was performed. Dual-isotope single photon emission computed tomographic/computed tomographic imaging was applied with the use of 123I to follow donor cell survival and distribution and with the use of 99mTC-tetrofosmin for perfusion imaging. In vitro, iodide uptake in NISpos-hiPSCs was increased 100-fold above that of nontransgenic controls. In vivo, viable NISpos-hiPSCs could be visualized for up to 15 weeks. Immunohistochemistry demonstrated that hiPSC-derived endothelial cells contributed to vascularization. Up to 12 to 15 weeks after transplantation, no teratomas were ...
TY - JOUR. T1 - A Drug Screen using Human iPSC-Derived Hepatocyte-like Cells Reveals Cardiac Glycosides as a Potential Treatment for Hypercholesterolemia. AU - Cayo, Max A.. AU - Mallanna, Sunil K.. AU - Di Furio, Francesca. AU - Jing, Ran. AU - Tolliver, Lauren B.. AU - Bures, Matthew. AU - Urick, Amanda. AU - Noto, Fallon K.. AU - Pashos, Evanthia E.. AU - Greseth, Matthew D.. AU - Czarnecki, Maciej. AU - Traktman, Paula. AU - Yang, Wenli. AU - Morrisey, Edward E.. AU - Grompe, Markus. AU - Rader, Daniel J.. AU - Duncan, Stephen A.. PY - 2017/4/6. Y1 - 2017/4/6. N2 - Efforts to identify pharmaceuticals to treat heritable metabolic liver diseases have been hampered by the lack of models. However, cells with hepatocyte characteristics can be produced from induced pluripotent stem cells (iPSCs). Here, we have used hepatocyte-like cells generated from homozygous familial hypercholesterolemia (hoFH) iPSCs to identify drugs that can potentially be repurposed to lower serum LDL-C. We found that ...
The general functions of integrins in attachment, gene expression, motility, polarity, shape, proliferation, and survival are well known. These critical functions provide reason for their wide expression across cell types. In particular, integrins are expressed in varying stem cell populations as well as other cell populations throughout the human body. Integrin alpha-6 (ITGA6) is a particular isoform of the integrin family, which is expressed across stem cell populations and has been shown to play an integral role in embryonic pluripotent stem cell self-renewal (Villa-Diaz 2016). Human embryonic stem cells (hESC) and human induced pluripotent stem cells (hiPSC) share the same morphology and gene expression but they do not share the same origin, since hESCs are obtained from the inner cell mass of blastocyst embryos, while hiPSCs are derived from somatic cells via genetic reprogramming. This similarity is what makes hiPSCs so widely applicable to regenerative medicine as they are more ethically ...
Human iPSC-Derived Neural Stem Cells from a healthy male donor. Axol iPSC-Derived Neural Stem Cells can be differentiated to neurons and glial cells.
The generation of human induced pluripotent stem cells (hiPSCs) with a high differentiation potential provided a new source for hepatocyte generation not only for drug discovery and in vitro disease models, but also for cell replacement therapy. However, the reported hiPSC-derived hepatocyte-like cells (HLCs) were not well characterized and their transplantation, as the most promising clue of cell function was not reported. Here, we performed a growth factor-mediated differentiation of functional HLCs from hiPSCs and evaluated their potential for recovery of a carbon tetrachloride (CCl)-injured mouse liver following transplantation. The hiPSC-derived hepatic lineage cells expressed hepatocyte-specific markers, showed glycogen and lipid storage activity, secretion of albumin (ALB), alpha-fetoprotein (AFP), urea, and CYP450 metabolic activity in addition to low-density lipoprotein (LDL) and indocyanin green (ICG) uptake. Similar results were observed with human embryonic stem cell (hESC)-derived ...
Reprogramming of somatic cells is a valuable tool to understand the mechanisms of regaining pluripotency and further opens up the possibility of generating patient-specific pluripotent stem cells. Reprogramming of mouse and human somatic cells into pluripotent stem cells, designated as induced pluri …
The Induced Pluripotent Stem Cell (iPSC) Repository is a major effort from CIRM to create a collection of stem cells developed from thousands of individuals. CIRM is creating the iPSC bank so that scientists can use the cells, either in a petri dish or transplanted into animals, to study how disease develops and progresses and develop and test new drugs or other therapies. The iPSC bank is now open and cell lines are available at catalog.coriell.org/CIRM. The large size of the collection will provide researchers with a powerful tool for studying genetic variation between individuals, helping scientists understand how disease and treatment may vary in a diverse population like Californias. What is the iPSCRepository? How does it work? Why iPS cells? Who is generating the cells? Which diseases will be represented? How many samples are being collected for each condition? What is the iPSCRepository? The Human Induced Pluripotent Stem Cell (hiPSC) Repositoryis one of the California stem cell agencys ...
A core area of our research in the field of biodistribution of cell therapies is immunohistochemical detection of hematopoietic stem cells and induced pluripotent stem cells in tissues and whole-organ slices of laboratory animals, even if the number of cells distributed throughout the body is low. For this purpose, a broad spectrum of antibodies is established at Fraunhofer ITEM. In view of safety aspects and registration, histopathology and immunohistochemistry are performed under GLP conditions.. A risk of stem cell therapies is the formation of teratomas. These germ cell tumors can be induced by injection of pluripotent stem cells into immunodeficient mice. During tumor growth, the injected stem cells differentiate into cell types from all three germ layers, reflecting embryonic development to a certain degree. With the broad range of methods we have at our disposal, teratoma formation can be assessed as an endpoint in toxicology studies. ...
by Monya Baker. Rat pluripotent stem cells could bring knock-out rats, reprogramming insights, and a larger menagerie of stem cells.. A quartet of papers in December describe the production of rat pluripotent stem cells, both from rat embryos and from the genetic manipulation of cultured rat cells.. The rat embryonic stem cells were derived by research teams led by Austin Smith in Cambridge, UK and Qi-Long Ying of the University of Southern California, Los Angeles. They mark the first of many attempts to create ES cells that, when mixed with a normal rat embryo, can contribute to the germline in the resultant rats. Though neither team has yet produced a knockin or knockout rat, Ying believes this could happen in less than a year. The key to success was finding a combination of small molecules capable of inhibiting differentiation, which Ying and Smith described in mouse embryonic stem cells in May. While pathways to promote self-renewal in stem cells may ...
[Generation of Zone-specific Hepatocyte-like Cells from Human Induced Pluripotent Stem Cells for Accurate Prediction of Drug-induced Hepatotoxicity]. Yakugaku Zasshi. 2019;139(12):1509-1512 Authors: Mitani S Abstract Human induced pluripotent stem (iPS) cell-derived hepatocyte-like cells (iPS-HLCs) are expected to be applicable to large-scale in vitro hepatotoxicity screening...
Insulin resistance leads to a number of metabolic and cellular abnormalities including endothelial dysfunction that increase the risk of vascular disease. Although it has been particularly challenging to study the genetic determinants that predispose to abnormal function of the endothelium in insulin-resistant states, the possibility of deriving endothelial cells from induced pluripotent stem cells generated from individuals with detailed clinical phenotyping, including accurate measurements of insulin resistance accompanied by multilevel omic data (eg, genetic and genomic characterization), has opened new avenues to study this relationship. Unfortunately, several technical barriers have hampered these efforts. In the present review, we summarize the current status of induced pluripotent stem cell-derived endothelial cells for modeling endothelial dysfunction associated with insulin resistance and discuss the challenges to overcoming these limitations. ...
0007] A currently known method for purifying cardiomyocytes is by preliminarily introducing a certain marker gene (e.g. GFP) into the genome of a stem cell (Non-Patent Document 1). However, this method requires genomic alteration, which itself presents an aesthetic problem and it also involves unpredictable serious risks in safety, such as a change in cells canceration rate (Non-Patent Document 2). A method involving genomic alteration has also been reported as a way to positively remove undifferentiated pluripotent stem cells (Non-Patent Document 3). A method taking a different approach has been reported, in which ceramide analogues known to have a cell death inducing action are used to induce cell death in embryonic stem cells in a comparatively specific way (Non-Patent Document 4). However, this method does not assure satisfactory removal of pluripotent stem cells since the group of cells cultured after treatment with the ceramide analogues contained (OCT positive cells) in an amount as much ...
The research published in the journal Nature Biotechnology shed more light on which genetic factors are critical in the reprogramming of adult skin cells to become other types of cell. In this study the researchers achieved reprogramming of adult cells without the use of a gene which has been linked to the development of tumours.. Prof Robin Lovell-Badge, Head of Developmental Genetics, MRC National Institute For Medical Research, said:. "This is a very interesting follow up study from Yamanaka lab, where the main conclusion is that, of the four factors they originally used to reprogramme either mouse or human skin cells (fibroblasts) into pluripotent stem cells, the oncogene cMyc is dispensable. This is important because of their previous data showing that chimeric mice derived using iPS cells were prone to tumours, which would obviously compromise the use of human iPS cells in research and for therapies. Chimeric mice made with the "3-factor" mouse iPS cells did not develop obvious tumours, ...
We studied the level of spontaneous telomere dysfunction in Rattus norvegicus (Berkenhout, 1769) (Rodentia, Muridae) embryonic fibroblasts (rEFs) and in cultured in vitro rat pluripotent stem cells (rPSCs), embryonic stem cells (rESCs) and induced pluripotent stem cells (riPSCs), on early passages and after prolonged cultivation. Among studied cell lines, rESCs showed the lowest level of telomere dysfunction, while the riPSCs demonstrated an elevated level on early passages of cultivation. In cultivation, the frequency of dysfunctional telomeres has increased in all studied cell lines; this is particularly true for dysfunctional telomeres occurring in G1 stage in riPSCs. The obtained data are mainly discussed in the connection with the specific structure of the telomere regions and their influence on the differential DNA damage response in them.
Although adipose tissue is an expandable and readily attainable source of proliferating, multipotent stem cells, its potential for use in regenerative medicine has not been extensively explored. Here we report that adult human and mouse adipose-derived stem cells can be reprogrammed to induced pluripotent stem (iPS) cells with substantially higher efficiencies than those reported for human and mouse fibroblasts. Unexpectedly, both human and mouse iPS cells can be obtained in feeder-free conditions. We discovered that adipose-derived stem cells intrinsically express high levels of pluripotency factors such as basic FGF, TGFβ, fibronectin, and vitronectin and can serve as feeders for both autologous and heterologous pluripotent cells. These results demonstrate a great potential for adipose-derived cells in regenerative therapeutics and as a model for studying the molecular mechanisms of feeder-free iPS generation and maintenance. ...
TY - JOUR. T1 - Pluripotent stem cells and tolerance induction in organ transplantation. AU - Imberti, Barbara. AU - Monti, Manuela. AU - Casiraghi, Federica. PY - 2015/2/21. Y1 - 2015/2/21. N2 - PURPOSE OF REVIEW: Ongoing research is constantly looking for means to modulate the immune system for long-lasting engraftment of pluripotent stem cells (PSC) during stem cell-based therapies. This study reviews data on in-vitro and in-vivo immunogenicity of embryonic and induced-PSC and describes how their immunological properties can be harnessed for tolerance induction in organ transplantation. RECENT FINDINGS: Although PSC display immunomodulatory properties in vitro, they are capable of eliciting an immune response that leads to cell rejection when transplanted into immune-competent recipients. Nevertheless, long-term acceptance of PSC-derived cells/tissues in an allogeneic environment can be achieved using minimal host conditioning. Protocols for differentiating PSC towards haematopoietic stem ...
Induced pluripotent stem cells are special because theyre not made from embryos. Instead, they come from harvesting skin cells from people, then treating those cells with genes that reverse the cells life stage back to its stem cell state. That means scientists are able to make induced pluripotent stem cells from cells taken from a patients own body. The resulting cells should be well matched to the patients own genetics, although its possible the "induction" part of the process introduces genetic aberrations into the cells. ...
The human cultures described here satisfy the criteria used to define pluripotent stem cells. These include presentation of a series of markers commonly used to identify pluripotent stem cells, morphological similarity to mouse ES and EG cells, normal and stable karyotype maintained over at least 10 passages, and demonstrated ability to differentiate into a wide variety of cell types.. The histological profile of these human cells (AP+, SSEA-1+, SSEA-3+, SSEA-4+, TRA-1-60+, and TRA-1-81+) differs from undifferentiated human embryonal carcinoma (EC) and rhesus ES cells, which are SSEA-1 negative (15, 38). The fact that differentiation of the human EC line NTERA2 leads to increased expression of SSEA-1 may suggest that this marker is indicative of differentiation in the human PGC-derived cultures. However, NTERA2 differentiation is accompanied by the loss of the other markers (39, 40), which we do not observe. A second possibility is that SSEA-1 reactivity reflects an intrinsic difference between ...
CAMBRIDGE, MA and SEATTLE, WA, October 3, 2017-BlueRock Therapeutics and Universal Cells Inc. today announced that they have entered into a collaboration and license agreement to create induced pluripotent stem (iPS) cell lines useful for the manufacture of allogeneic cellular therapies.. "BlueRock is at the cutting edge of the cell therapy field and our collaboration with the company represents an important step in their efforts to develop off-the-shelf cell therapies for degenerative diseases," said Claudia Mitchell, Ph.D., CEO of Universal Cells. "We are honored that our technology will support the advancement of BlueRocks cell therapies.". Under the terms of the agreement, BlueRock and Universal Cells will collaborate to engineer iPS cell lines for clinical use. The agreement grants BlueRock commercial rights to use the engineered cell lines for undisclosed therapeutic applications.. "This partnership with Universal Cells provides us with an important technology useful to advance ...
Latest publications. Shinya Yamanaka, MD, PhD - Fujishiro SH, Nakano K, Mizukami Y, Azami T, Arai Y, Matsunari H, Ishino R, Nishimura T, Watanabe M, Abe T, Furukawa Y, Umeyama K, Yamanaka S, Ema M, Nagashima H, Hanazono Y. Generation of Naive-like Porcine Induced Pluripotent Stem Cells Capable of Contributing to Embryonic and Fetal Development. Stem Cells Dev. 2012 Aug 13; Shinya Yamanaka, MD, PhD - Blanpain C, Daley GQ, Hochedlinger K, Passegue E, Rossant J, Yamanaka S. (2012) Stem cells assessed. Nat Rev Mol Cell Biol 13: 471-6; Shinya Yamanaka, MD, PhD - Tanaka T, Takahashi K, Yamane M, Tomida S, Nakamura S, Oshima K, Niwa A, Nishikomori R, Kambe N, Hara H, Mitsuyama M, Morone N, Heuser JE, Yamamoto T, Watanabe A, Sato-Otsubo A, Ogawa S, Asaka I, Heike T, Yamanaka S, Nakahata T, Saito MK. Induced pluripotent stem cells from CINCA syndrome patients as a model for dissecting somatic mosaicism and drug discovery. Blood. 2012 Aug 9; 120(6):1299-308; Shinya Yamanaka, MD, PhD - Isobe H, Shoji M, ...
Establishment of porcine induced pluripotent stem cells Jenn-Rong Yang. The purpose of this study is to establish and study pluripotency, efficiency of embryoid body formation, and potency of differentiation of porcine induced pluripotent stem (piPS) cells. The results showed that the morphology of porcine ear fibroblasts were transformed into colony type from spindle type one month after infection with defined factors of Oct4, Sox2, Klf, and c-Myc. After co-cultured with STO cells, typical colony morphology of stem cells was found. The piPS cells expressed pluripotent cellular markers including Oct-4, AP, SSEA-3, SSEA-4, TRA-1-60, and TRA-1-81 antigens. The efficiency of embryoid body formation of pips cells was excellent when induced by using hanging drop method. The karyotype of the piPS cells was also normal (36+XY). In addition, transplantation of piPS cells into NOD-SCID mice resulted teratomas .Various differentiated cells were found in the teratoma, such as neural cells, keratinocytes, ...
The present disclosure provides isolated naïve pluripotent stem cells as well as methods and compositions of generating and culturing the same.
Methods : Fibroblasts from two L-ORD affected and two unaffected siblings were reprogrammed into induced pluripotent stem cells (iPSCs) using sendai viral vectors containing the four Yamanaka factors. iPSCs were fully characterized via immunocytochemistry, karyotyping, and spontaneous differentiation of embryoid bodies into all three germ layers. iPSCs were then differentiated into RPE using a developmentally guided protocol. Cell shapemetric analysis was performed using custom software on iPSC-derived RPE stained with ZO-1. Apical media was analyzed for protein and lipid composition. Results : Although iPSC-derived RPE from patients and unaffected siblings have similar hexagonality (score, side ratio, area ratio), L-ORD patient iPSC-derived RPE tended be more variable in size and on average possessed greater cell areas. Mass Spect analysis revealed elevated sequence coverage of glutathione, cystatin C, keratins, vimentin, actin, and tubulin secreted by patient iPSC-RPE. Lipidomic analysis of ...
Video articles in JoVE about fibroblast growth factor 2 include High Efficiency Differentiation of Human Pluripotent Stem Cells to Cardiomyocytes and Characterization by Flow Cytometry, An Optogenetic Approach for Assessing Formation of Neuronal Connections in a Co-culture System, Directed Differentiation of Primitive and Definitive Hematopoietic Progenitors from Human Pluripotent Stem Cells, An Enzyme- and Serum-free Neural Stem Cell Culture Model for EMT Investigation Suited for Drug Discovery, Culturing Human Pluripotent and Neural Stem Cells in an Enclosed Cell Culture System for Basic and Preclinical Research, A Unified Methodological Framework for Vestibular Schwannoma Research, Defined Xeno-free and Feeder-free Culture Conditions for the Generation of Human iPSC-derived Retinal Cell Models, Preclinical Model of Hind Limb Ischemia in Diabetic Rabbits, Using Human Induced Pluripotent Stem Cell-derived Hepatocyte-like Cells for Drug Discovery, A cGMP-applicable Expansion Method
To date, human primary islets have been considered the gold standard for diabetes research as they provide a model to assess key criteria such as insulin production, insulin secretion, and in vitro toxicity. However, availability of these cells can be limiting. Moreover, primary islets can be unreliable: they show large donor variation, preventing the generation of consistent results.. Beta cells derived from human induced pluripotent stem cells are a powerful alternative to primary islets, as they offer a virtually unlimited source of cells, are easy to culture, and provide reliable and reproducible models for drug discovery and toxicity testing.. Cellartis hiPSC-derived beta cells provide a reliable, off-the-shelf source of beta cells. These cells consistently demonstrate high expression of known beta cell mRNA transcripts and proteins, and they display appropriate responsiveness to incretin and KCl stimulation. Our hiPSC-derived beta cells are an excellent tool for studying the differences ...
Abstract Human stem cell research is a new field with much promise, but progress towards a clinical setting has been complicated by scientific and ethical challenges. The most heated discussion over stem cell research has focused on the source of human embryonic stem cells (ESCs). Different views on the moral status of the human embryo have plagued all aspects of the debate (and decision-making). In 2006, a way of de-differentiating somatic cells to a pluripotent state was realised. The advent of these induced pluripotent stem cells (iPSCs) appeared to circumvent concerns over embryo destruction, and hence iPSCs have been touted as an ethical way forward. However, for the foreseeable future, scientific investigations involving iPSCs are likely to drive further embryo destruction. As a result, iPSC research is complicit in embryo destruction and is inextricably locked in to the moral status debate. I argue that a new approach is needed to deal with the serious uncertainties and indeterminate ...
Murine pluripotent stem cells can exist in two distinct states, blastocyst-derived LIF-dependent embryonic stem cells (ESCs) and epiblast-derived bFGF-dependent stem cells (EpiSCs). Murine ESCs and similar iPSC lines are more of the "ground-state" in terms of developmental status, as reflected by the lack of X chromosome inactivation in female cells and their abilities to pass as single cells. Human iPSCs, like human ES cells, are more similar to mouse EpiSCs. Unfortunately these human pluripotent stem cells are difficult to genetically manipulate, e.g. knockin or knockout. They also grow slowly, with doubling time averaging 36 hours. In order to create ground-state human iPSCs, several approaches have been tested, including reprogramming iPSC-derived fibroblasts, continuously expressing 5 iPS factors (Oct4, Sox2, Nanog, c-Myc, and Klf4), or using chemicals to inhibit chromatin modifying enzyme HDAC. While these approaches succeeded to certain degrees, the resulting cell lines seem to have some ...
Another objection is the iPSCs are just like embryonic stem cells and so are no good for treating patients. It is true that because iPSCs are pluripotent like embryonic stem cells they will likely have many of the same safety issues for transplantation. More research is needed. What iPSCs are good for is creating model tissues for scientists to use to study disease progression and treatment. As I have written before, previous to iPSCs, scientists would have to create a mouse or other animal that exhibited the symptoms of a human disease that they were interested in studying. Now they can take a skin cell from a person with a disease, reprogram that cell back to a pluripotent state, and then differentiate them into cells of interest whether they be neurons or fat cells. iPSCs can continue to grow in culture and be frozen giving researchers a nearly limitless supply of diseased cells to work on. This is especially useful in brain disorders because isolating neurons from the brain of a patient is ...
ATCC provides numerous examples of human inducible pluripotent stem cell lines (iPSC) designed to support physiologically relevant, in vitro research in regenerative medicine.
ATCC provides numerous examples of human inducible pluripotent stem cell lines (iPSC) designed to support physiologically relevant, in vitro research in regenerative medicine.
Recent progresses in the field of Induced Pluripotent Stem Cells (iPSCs) have opened up many gateways for the research in therapeutics. iPSCs are the cells which are reprogrammed from somatic cells using different transcription factors. IPSCs possess unique properties of self renewal and differentiation to many types of cell lineage. Hence could replace the use of embryonic stem cells, and may overcome the various ethical issues regarding the use of embryos in research and clinics. Overwhelming responses prompted worldwide by a large number of researchers about the use of iPSCs evoked a large number of peple to establish more authentic methods for iPSC generation. This would require understanding the underlying mechanism in a detailed manner. There have been a large number of reports showing potential role of different molecules as putative regulators of iPSC generating methods. The molecular mechanisms that play role in reprogramming to generate iPSCs from different types of somatic cell sources
Somatic cells could be reprogrammed to induced pluripotent stem cells (iPS) by ectopic expression of OCT4, SOX2, KLF4 and MYC (OSKM). We aimed to gain insights into the early mechanisms underlying the induction of pluripotency. GSE28688 containing 14 gene expression profiles were downloaded from GEO, including untreated human neonatal foreskin fibroblasts (HFF1) as control, OSKM-induced HFF1 (at 24, 48, 72 h post-transduction of OSKM encoding viruses), two iPS cell lines, and two embryonic stem (ES) cell lines. Differentially expressed genes (DEGs) were screened between different cell lines and the control by Limma package in Bioconductor. KEGG pathway enrichment analysis was performed by DAVID. The STRING database was used to construct protein-protein interaction (PPI) network. Activities and regulatory networks of transcription factors (TFs) were calculated and constructed by Fast Network Component Analysis (FastNCA). Compared with untreated HFF1, 117, 347, 557, 2263 and 2307 DEGs were obtained from
The STEMdiff™ Pancreatic Progenitor Kit was optimized with reproducibility in mind. We designed both the medium formulation as well as the protocol to maximize the efficiency of differentiation across multiple human embryonic stem (ES) and induced pluripotent stem (iPS) cell lines (see Figure 2 here for the supporting data). At STEMCELL Technologies, we routinely use the Wisconsin H1 (WA1) and H9 (WA9) human ES cell lines during the development of our STEMdiff™ products. In addition, we have acquired or generated many iPS cell lines, derived from a variety of somatic cell types. Results generated using these cell lines indicate that the user of this kit can expect to see on average, greater than 65% of cells co-expressing PDX-1 and NKX6.1 at the end of Stage 4. Some experiments have generated greater than 90% pure populations of pancreatic progenitors. We do not see a significant difference in the efficiency of differentiation between human ES and iPS cell lines. Within a given experiment, ...
Induced pluripotent stem cells (iPS cells) are important for the future development of regenerative medicine involving autologous cell therapy. Before autologous cell therapy can be applied to human patients, suitable animal models must be developed, and in this context non-human primate models are critical. We previously characterized several lines of marmoset iPS cells derived from newborn skin fibroblasts. In the present studies, we explored methods for the directed differentiation of marmoset iPS cells in the neuroectodermal lineage. In this process we used an iterative process in which combinations of small molecules and protein factors were tested for their effects on mRNA levels of genes that are markers for the neuroectodermal lineage. This iterative process identified combinations of chemicals/factors that substantially improved the degree of marker gene expression over the initially tested combinations. This approach should be generally valuable in the directed differentiation of ...