人胚胎干细胞和诱导性多功能干细胞饲养层细胞的制备方法  

Derivation and preparation of feeder cells for culturing human embryonic and induced pluripotent stem cells

作　　者：谭小兵[1] George T-J Huang 

机构地区：[1]云南省第一人民医院口腔内科,昆明650032 [2]美国波士顿大学牙学院牙体牙髓病学系,波士顿ma02118

出　　处：《国际口腔医学杂志》2014年第3期268-271,共4页International Journal of Stomatology

基　　金：国家自然科学基金地区科学基金项目(81360161)

摘　　要：目的建立一种能维持人胚胎干细胞（ESC）和诱导性多功能干细胞（iPSC）生长的饲养层细胞的标准制备方法。方法选取受孕12．5-14．5d的CF-1雌鼠，原代培养的鼠胚胎成纤维细胞（MEF）经丝裂霉素C处理后作为饲养层细胞，观察其维持人ESC和iPSC的生长情况；同时研究不同培养器具所需细胞数量，制备高质量饲养层细胞。结果经丝裂霉素C处理后，MEF可很好维持人ESC和iPSC的生长，使其保持自我更新能力和未分化状态；控制铺板的细胞密度，可得到高质量饲养层细胞。结论本实验得到的饲养层细胞能很好支持人ESC和iPSC生长，可作为饲养层细胞制备的标准方法使用。Objective This study aimed to establish a standard method to derive and prepare feeder cells for the optimized growth of human embryonic stem cells（ESCs） and induced pluripotent stem cells（iPSCs）. Methods CF-1 mouse fetuses between 12.5 and 14.5 days of gestation were separated, trypsinized, and cultured to derive primary mouse embryonic fibroblasts（MEFs）. MEFs were then treated with mitomycin C to prepare feeder cells. The number of cells needed for high-quality feeder cells was determined through a variety of cell culture vessels. Results MEF from CF-1 pregnant mouse treated with mitomycin C could be used as feeder cells to support the culture and to maintain the undifferentiating of ESCs and iPSCs. High-quality feeder cells could be derived if an appropriate number of cells were plated. Conclusion MEF prepared in this method can optimally support and maintain the undifferentiated human ESCs and iPSCs. Furthermore, this method can be utilized as a standard method to derive feeder cells.

关 键 词：人胚胎干细胞 人诱导性多功能干细胞 饲养层细胞 鼠胚胎成纤维细胞 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]