Yang Ke, Dong Juan, Xu Lan, Zhou Zhenning, Wang Qin, Ding Xiaoyan
School of Life Sciences and Technology, Shanghai Jiaotong University, Shanghai 200240, China.
Sheng Wu Gong Cheng Xue Bao. 2008 Oct;24(10):1783-9.
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.
未分化的胚胎干细胞（ES细胞）如果在细胞因子白血病抑制因子（LIF）存在的情况下进行培养，就能够在体外维持。ES细胞也能够在体外分化。一种特别有效的诱导ES细胞分化的方法是在没有LIF的情况下将ES细胞培养成聚集体。在这些条件下，它们会形成被称为胚状体（EBs）的结构。然而，目前形成EB的方案仍然多种多样。为了便于进一步研究，我们仔细控制了EB形成的培养条件，在此我们报告一种有效的方案，通过该方案获得了均匀分化的EB，并通过测量跳动心肌细胞的分化进行监测。此外，通过使用该方案，我们观察到在长期培养（>60天）的接种EB中仍然存在具有多能性的细胞集落。这一观察结果提出了一种潜在的可能性，即ES细胞可能在由分化细胞提供的微环境中保持多能性。
