长期培养小鼠胚胎干细胞拟胚体(EB)的观察

Persistently Exist of ES-like Cell Population in Long-term Cultured Embryoid Bodies

摘要:

胚胎干细胞在体外培养条件下能够维持自我更新, 并具有向多种细胞类型分化的能力, 因此被广泛用于研究细胞分化的分子机理以及药物筛选。形成拟胚体(Embryoid body, EB)是胚胎干细胞分化常用的技术手段。为了便于今后利用EB做进一步的药物筛选及分化研究, 我们严格规范了形成EB的条件, 得到了分化状态均一性很高的EB。利用这一条件, 我们观察到在分化条件下长期培养(长达60 d)的EB中仍有表达各项多能性指标的细胞集落。有关这一现象的进一步分析工作正在进行中。

Abstract:

Undifferentiated embryonic stem (ES) cells can be maintained in vitro if cultured in the presence of the cytokine leukaemia inhibitory factor (LIF). ES cells can also differentiate in vitro. A particularly efficient method for inducing ES cell differentiation is to culture ES cells as aggregates in the absence of LIF. Under these conditions they form structures known as embryoid bodies (EBs). However the current protocols for EB formation are still diverse. In order to facilitate further study, we carefully controlled the culture conditions for EB formation, and here we report an efficient protocol by which uniformly differentiated EBs were obtained, monitored by measuring the differentiation of beating cardiomyocytes. Furthermore, by using this protocol we observed in long-term cultured plating EBs (>60 days) there still exist cell colony with pluripotency. This observation raised a potential possibility that ES cells may keep pluripotent in a niche provided by differentiated cells.