The Art of Capturing Pluripotency: Creating the Right Culture.

Qi-Long Ying,Austin Smith

doi:10.1016/j.stemcr.2017.05.020

Abstract

Embryonic stem cells (ESCs) are a unique tool for genetic perturbation of mammalian cellular and organismal processes additionally in humans offer unprecedented opportunities for disease modeling and cell therapy. Furthermore, ESCs are a powerful system for exploring the fundamental biology of pluripotency. Indeed understanding the control of self-renewal and differentiation is key to realizing the potential of ESCs. Building on previous observations, we found that mouse ESCs can be derived and maintained with high efficiency through insulation from differentiation cues combined with consolidation of an innate cell proliferation program. This finding of a pluripotent ground state has led to conceptual and practical advances, including the establishment of germline-competent ESCs from recalcitrant mouse strains and for the first time from the rat. Here, we summarize historical and recent progress in defining the signaling environment that supports self-renewal. We compare the contrasting requirements of two types of pluripotent stem cell, naive ESCs and primed post-implantation epiblast stem cells (EpiSCs), and consider the outstanding challenge of generating naive pluripotent stem cells from different mammals.

Highlights

Embryonic stem cells (ESCs) were originally derived by co-culture with a feeder layer of mitotically inactivated fibroblasts in medium containing fetal calf serum
It was found that feeders could be replaced by the cytokine leukemia inhibitory factor (LIF) (Smith et al, 1988; Smith and Hooper, 1987; Williams et al, 1988) and that serum could be substituted by bone morphogenetic protein (BMP) (Ying et al, 2003)
Using titrated CHIR we found that partial inhibition of glycogen synthase kinase 3 (GSK3) had a short-term stimulatory effect on ESC self-renewal in the absence of LIF and serum

Summary

Introduction

ESCs were originally derived by co-culture with a feeder layer of mitotically inactivated fibroblasts in medium containing fetal calf serum. LIF supports ESC self-renewal through receptor-mediated stimulation of Janus-associated kinase (JAK) and activation of the transcription factor Stat3 (Niwa et al, 1998). Using titrated CHIR we found that partial inhibition of GSK3 had a short-term stimulatory effect on ESC self-renewal in the absence of LIF and serum.

Results

Conclusion