원문정보
초록
영어
Human pluripotent stem cells (hPSCs) self-renew indefinitely as highly organized pluripotent colonies; this self-renewal is evident for both human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs). Unlike mouse pluripotent stem cell colonies, human colonies form a uniform, flat epithelium-like monolayer. Interestingly, it has been reported that colony morphology is closely correlated with the maintenance of pluripotency. However, the molecular mechanisms that underlie human pluripotent colony formation and organization are poorly understood. In this study, using real-time imaging tools, we examine in vitro colony formation by enzymatically dissociated single hPSCs (specifically CHA15 cells, H9 hESCs and hiPSCs) under feeder-free conditions. We showed that colony formation consists of 4 stages: attachment, migration, colony formation (aggregation), and colony organization, which are facilitated in an intracellular calcium-dependent manner. Moreover, we found that blocking Gi-coupled GPCR signaling results in enhanced cell-cell interactions and plays an integral role in promoting the survival of hESCs in culture. Using various visualization methods, we identified conditions required for colony formation, and we suggested a promising new target for modulating hPSC colony formation and organization. These results are likely to be useful for engineering hPSCs to further study the mechanisms involved in pluripotency.