Abstract
SummaryEmbryonic stem cells (ESCs) of mice and humans have distinct molecular and biological characteristics, raising the question of whether an earlier, “naive” state of pluripotency may exist in humans. Here we took a systematic approach to identify small molecules that support self-renewal of naive human ESCs based on maintenance of endogenous OCT4 distal enhancer activity, a molecular signature of ground state pluripotency. Iterative chemical screening identified a combination of five kinase inhibitors that induces and maintains OCT4 distal enhancer activity when applied directly to conventional human ESCs. These inhibitors generate human pluripotent cells in which transcription factors associated with the ground state of pluripotency are highly upregulated and bivalent chromatin domains are depleted. Comparison with previously reported naive human ESCs indicates that our conditions capture a distinct pluripotent state in humans that closely resembles that of mouse ESCs. This study presents a framework for defining the culture requirements of naive human pluripotent cells.
Highlights
Human pluripotent stem cells, including embryonic stem cells (ESCs) and induced pluripotent stem cells, share molecular and functional properties with epiblast stem cells (EpiSCs) derived from the mouse postimplantation epiblast (Brons et al, 2007; Tesar et al, 2007)
We took a systematic approach to identify small molecules that support self-renewal of naive human Embryonic stem cells (ESCs) based on maintenance of endogenous OCT4 distal enhancer activity, a molecular signature of ground state pluripotency
Iterative chemical screening identified a combination of five kinase inhibitors that induces and maintains OCT4 distal enhancer activity when applied directly to conventional human ESCs
Summary
Human pluripotent stem cells, including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), share molecular and functional properties with epiblast stem cells (EpiSCs) derived from the mouse postimplantation epiblast (Brons et al, 2007; Tesar et al, 2007). Ng and colleagues screened a combination of 20 compounds for enhanced expression of NANOG in mTesr, a customized medium for human ESCs containing high levels of FGF and TGFb. This study reported that a combination of 2i, hLIF, and Dorsomorphin induced upregulation of a number of genes expressed in the human preimplantation embryo (Chan et al, 2013). This study reported that a combination of 2i, hLIF, and Dorsomorphin induced upregulation of a number of genes expressed in the human preimplantation embryo (Chan et al, 2013) In contrast with these two studies, two other recent papers reported that 2i and FGF are sufficient to maintain naive-like human ESCs in the presence (Valamehr et al, 2014) or absence (Ware et al, 2014) of hLIF
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
