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Cell Journal، جلد ۱۶، شماره Suppl ۱، صفحات ۱۱۸-۱۱۸
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عنوان انگلیسی Ps-87: A Novel, Safe Strategy to Derive Naïve-Like Human Pluripotent Stem Cells
چکیده انگلیسی مقاله Objective: Although human and mouse embryonic stem cells (ESCs) are both derived from the inner cell mass of the blastocysts, they exhibit distinct characteristics particularly in terms of developmental potential. While mouse ESCs exist in a naïve pluripotent state, human ESCs are in a primed state, displaying a propensity to better differentiate into certain cell types than others. Induction of naïve pluripotency in the human pluripotent stem cells (PSCs) is thus of particular importance to obtain pluripotent cells with less differentiation propensity. Here we report a novel safe approach to derive naïve-like human PSCs. Materials and Methods: Primed human ESCs and induced pluripotent stem cells (iPSCs) were first cultured in a mouse ESC culture medium containing LIF and the 2i inhibitors: PD0325901 which inhibits FGF signaling and CHIR99021 which blocks the GSK3 pathway. Next the cells were treated for five to seven days with specific small molecules. Once emerged, the naïve-like colonies were trypsinized and plated onto feeder-coated plates for expansion. Finally, naïve-like cells were characterized for properties of naïve pluripotency. Results: Our small-molecule cocktail could successfully induce naïve-like human PSCs which exhibited the naïve characteristics in terms of morphology, expandability, doubling time, single cell survival, single cell clonogenicity, LIF responsiveness, XIST repression, in vitro and in vivo differentiation, enhanced NANOG and E-Cadherin expression, and nuclear localization of the proteins STAT3 and TFE3. Conclusion: We have successfully devised a novel culture system for the safe derivation of naïve-like human PSCs from primed human PSCs using the combination of two chemicals under the 2i+LIF culture condition.
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نشانی اینترنتی http://celljournal.org/journal/article/abstract/412
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