【作者】 乌云她娜； 塔娜； 于海泉；

【Author】 Uyutana;Tana;Yu Hai-quan;Research Center for Laboratory Animal Science,Inner Mongolia University;

【机构】 内蒙古大学生命科学学院实验动物研究中心；

【摘要】 目的近年来研究发现,通过PD0325901和CHIR99021两种小分子抑制剂(2i)分别抑制FGF信号通路的丝裂原活化蛋白激酶和WNT信号通路的糖原合酶激酶3,显著促进了小鼠及大鼠干细胞多能性的维持。另外研究表明,基因组DNA甲基化与去甲基化等表观遗传修饰对体细胞去分化为干细胞,或干细胞再分化为体细胞极为关键。目前牛胚胎干细胞还没有被成功分离获得,2i等培养体系是否适合牛胚胎干细胞(bESC)的多能性维持及其作用机制仍不清楚。方法本研究通过在人胚胎干细胞培养基mTeSR1基础上添加PD0325901和CHIR99021两种小分子抑制剂(2i)和DNA甲基化转移酶抑制剂5-AzaC建立(3i)培养体系分离和鉴定牛类胚胎干细胞,研究药物处理后对牛胚胎干细胞的多能性维持及多能因子的表达的影响,结果3i培养体系有利于bESCs细胞增殖、存活和多能性相关基因的表达,本研究为牛胚胎干细胞系的建立提供了研究基础。更多还原

【Abstract】 Recently,it was found that the pluripotency of stem cells in mice and rats,were significantly improved by using the double inhibition(2 i) culture system,in which mitogenactivated protein kinase kinase(MAP2 K) in the fibroblast growth factor signaling(FGF) pathway,and glycogen synthase kinase 3(GSK3) in the WNT pathway were inhibited,other studies have shown that epigenetic modification,such as genomic DNA methylation and demethylation were critical to dynamic change of cell differentiation.To date,bovine embryonic stem cells(bESCs)have not been successfully isolated,it is not clear if the 2 i culture system are suitable for the maintain of bESCs pluripotency.This research was designed to study the effect of two small molecule inhibitors(2 i),PD0325901 and CHIR99021,combined with DNA methylation transferase inhibitor5-AzaC on the maintain of bESCs pluripotency and the expression of related molecular,which system based on the human embryonic stem cells culture medium mTeSRl our results showed that this 3 i culture system were beneficial to the maintain of bESCs pluripotency.更多还原