饲养层细胞体系及白血病抑制因子对维持小鼠胚胎干细胞未分化状态的作用  

Effect of Feeder Layer Culture System and Leukemia Inhibitory Factor on Keeping Mouse Embryonic Stem Cells in Undifferentiation

作　　者：赵政凯 刘畅[2] 刘福强[2] 高巍[3] 任甫[3] 刘力平 

机构地区：[1]辽宁省金秋医院神经内科,辽宁沈阳110000 [2]辽宁医学院附属第一医院内分泌科 [3]辽宁医学院解剖教研室,辽宁锦州121000

出　　处：《解剖科学进展》2009年第3期283-285,289,共4页Progress of Anatomical Sciences

基　　金：辽宁省自然基金资助(No.20052129);辽宁省科技攻关计划项目资助(No.2006225001-14)

摘　　要：目的建立并筛选最适合的小鼠胚胎干(ES)细胞饲养层培养体系。方法建立7种经过不同处理的鼠胚成纤维细胞作为饲养层培养体系,分别进行小鼠胚胎干细胞的培养,观察7种饲养层培养体系对小鼠ES细胞增殖能力及未分化状态的维持情况。结果小鼠胚胎细胞在3代内均可用于ES细胞的培养。与其他各组比较,第3代培养鼠胚成纤维细胞经丝裂霉素C处理2h后加外源白血病抑制因子(IF,10μg/ml)用作饲养层的克隆形成率和碱性磷酸酶(AKP)染色阳性度最高。结论外源性白血病抑制因子是抑制小鼠ES细胞分化的主要因素。Objective To study the effect of feeder layer culture system and leukemia inhibitory factor on keeping mice embryonic stem cells in undifferentiation. Methods Mice embryonic stem cells were isolated and cultured using mice embryonic fibroblasts as 7 different kinds of feeder layers, and the effects of different kinds of feeder layers on the development of mice embryonic stem cells were observed. Results 1-3 generations of mice embryonic cells were cultured into embryonic stem cells. The colony formation rate and degree of alkaline phosphatase （AKP） positive staining were the highest in the third generation of cultured mouse embryo fibroblast treated with mitomycin C for 2h as the feeder layer ＋ LIF10 μg/ml, compared with the other six kinds of feeder layers. Conclusion Ectogenic leukemia inhibitory factor is the prime factor inhibiting the differentiation of mice embryonic stem cells.

关 键 词：胚胎干细胞 饲养层 小鼠 白血病抑制因子 

分 类 号：Q813.11[生物学—生物工程]