Stably transfected human embryonic stem cell clones express OCT4-specific green fluorescent protein and maintain self-renewal and pluripotency
TLDR
These OCT4‐EGFP clonal cell lines exhibit features similar to parental hESCs, are pluripotent, and are able to produce all three embryonic germ layer cells, and they will be invaluable for studying not only OCT4 function in hESC self‐renewal and differentiation but also the factors required for maintenance of undifferentiated h ESCs in culture.Abstract:
Human embryonic stem cells (hESCs) are derived from the inner cell mass of preimplantation embryos; they can be cultured indefinitely and differentiated into many cell types in vitro. These cells therefore have the ability to provide insights into human disease and provide a potential unlimited supply of cells for cell-based therapy. Little is known about the factors that are important for maintaining undifferentiated hESCs in vitro, however. As a tool to investigate these factors, transfected hES clonal cell lines were generated; these lines are able to express the enhanced green fluorescent protein (EGFP) reporter gene under control of the OCT4 promoter. OCT4 is an important marker of the undifferentiated state and a central regulator of pluripotency in ES cells. These OCT4-EGFP clonal cell lines exhibit features similar to parental hESCs, are pluripotent, and are able to produce all three embryonic germ layer cells. Expression of OCT4-EGFP is colocalized with endogenous OCT4, as well as the hESC surface antigens SSEA4 and Tra-1-60. In addition, the expression is retained in culture for an extensive period of time. Differentiation of these cells toward the neural lineage and targeted knockdown of endogenous OCT4 expression by RNA interference downregulated the EGFP expression in these cell lines, and this correlates closely with the reduction of endogenous OCT4 expression. Therefore, these cell lines provide an easy and noninvasive method to monitor expression of OCT4 in hESCs, and they will be invaluable for studying not only OCT4 function in hESC self-renewal and differentiation but also the factors required for maintenance of undifferentiated hESCs in culture.read more
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HIF Induces Human Embryonic Stem Cell Markers in Cancer Cells
Julie Mathieu,Zhan Zhang,Wenyu Zhou,Amy Wang,John M. Heddleston,Claudia M A Pinna,Alexis Hubaud,Bradford M. Stadler,Michael Choi,Merav Bar,Muneesh Tewari,Alvin Y. Liu,Robert L. Vessella,Robert C. Rostomily,Donald E. Born,Marshall S. Horwitz,Carol B. Ware,C. Anthony Blau,Michele A. Cleary,Jeremy N. Rich,Hannele Ruohola-Baker +20 more
TL;DR: Hypoxia, through hypoxia-inducible factor (HIF), can induce an hESC-like transcriptional program, including the induced pluripotent stem cell (iPSC) inducers, OCT4, NANOG, SOX2, KLF4, cMYC, and microRNA-302 in 11 cancer cell lines.
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Efficient Differentiation of Hepatocytes from Human Embryonic Stem Cells Exhibiting Markers Recapitulating Liver Development In Vivo
David C. Hay,Debiao Zhao,Judy Fletcher,Zoe Hewitt,Doris McLean,Alai Urruticoechea-Uriguen,James R. Black,Cliff Elcombe,James A. Ross,Roland Wolf,Wei Cui,Wei Cui +11 more
TL;DR: Evidence is provided that hESCs can be differentiated efficiently in vitro to functional hepatocytes, which may be useful as an in vitro system for toxicity screening in drug discovery and evidence that the hESC‐derived hepatocytes are able to carry out a range of hepatocyte functions.
Journal ArticleDOI
Differentiation of Human Embryonic Stem Cells to Neural Lineages in Adherent Culture by Blocking Bone Morphogenetic Protein Signaling
TL;DR: It is shown that inhibition of bone morphogenetic protein signaling by its antagonist noggin is sufficient to block extraembryonic cell fate, transiently sustain Oct4 gene expression, and result in robust production of neural progenitors.
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Identification and targeting of the ROSA26 locus in human embryonic stem cells
TL;DR: The identification of the human homolog of the mouse Rosa26 locus is reported, demonstrating targeting of a red-fluorescent protein (tdRFP) cDNA to this locus through homologous recombination and expression of this targeted reporter in multiple hES cell–derived lineages.
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Regulation of Apoptosis and Differentiation by p53 in Human Embryonic Stem Cells
Han Qin,Tianxin Yu,Tingting Qing,Yanxia Liu,Yang Zhao,Jun Cai,Jian Li,Zhihua Song,Xiuxia Qu,Peng Zhou,Jiong Wu,Mingxiao Ding,Hongkui Deng +12 more
TL;DR: It is demonstrated that p53 protein accumulates in apoptotic hESCs induced by agents that damage DNA, however, despite the accumulation of p53, it nevertheless fails to activate the transcription of its target genes.
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