Institut für Anatomie, Universität Duisburg-Essen, Universitätsklinikum, Hufelandstr. 55, 45122, Essen, Germany.
In Vitro Cell Dev Biol Anim. 2010 Jun;46(6):516-28. doi: 10.1007/s11626-010-9272-8. Epub 2010 Feb 5.
Colonies of rhesus monkey embryonic stem cells (rhESC; cell line R366.4) have been described before to show a spatially ordered process of epithelial-mesenchymal transition in vitro. In the present investigations, we have studied variables of culturing conditions which influence the reproducibility of the formation of crater-like ingression centers in the colonies. Critical parameters are found to be age and density of mouse embryonic fibroblast (MEF) feeder cell layers, the mode of mitotic inactivation of the MEFs (mitomycin C, or irradiation), and the mode of rhESC isolation during subculturing (enzymatic/mechanical cell cluster isolation; type of enzyme). The described culturing system appears to offer a reproducible in vitro model potentially useful for studies on cellular processes involved in gastrulation in the primate.
恒河猴胚胎干细胞（rhESC；细胞系 R366.4）的集落以前曾被描述为在体外表现出空间有序的上皮-间充质转化过程。在本研究中，我们研究了影响集落中 crater-like 内陷中心形成的可重复性的培养条件变量。发现关键参数是小鼠胚胎成纤维细胞（MEF）饲养细胞层的年龄和密度、MEF 有丝分裂失活的方式（丝裂霉素 C 或辐照），以及亚培养过程中 rhESC 分离的方式（酶/机械细胞团分离；酶的类型）。所描述的培养系统似乎提供了一个可重复的体外模型，可能对研究灵长类动物中涉及原肠胚形成的细胞过程有用。
