丝裂霉素C对小鼠胎儿成纤维细胞增殖抑制作用的研究  

The Effects of Fibroblasts Feeder Layer Treated with Mitomycin C in Different Times on Mouse Embryonic Stem Cell Culture

作　　者：袁龙[1] 郭庆勇[1] 武聚富[1] 张慧[1] 况玲[1] 

机构地区：[1]新疆农业大学动物医学学院,乌鲁木齐830052

出　　处：《动物医学进展》2007年第5期41-45,共5页Progress In Veterinary Medicine

摘　　要：为探讨丝裂霉素C对小鼠胎儿成纤维细胞增殖的抑制作用,用10μg/mL的丝裂霉素C处理昆明白胎鼠P1代成纤维细胞1.5、2、2.5、3、3.5、4 h,用台盼蓝和AO/PI染色法观察细胞死亡情况,MTT测其增殖情况,PCNA染色观察其增殖细胞状态,比较胚胎干细胞接种在冻存过的饲养层细胞与未冻存的饲养层细胞的生长情况。结果表明,丝裂霉素C处理MEF时间小于3 h,不能抑制其生长,处理4 h后,饲养层细胞部分已经没有核仁,部分细胞已经崩解,且胚胎干细胞接种在冻存过的饲养层细胞与未经冻存的饲养层细胞都生长良好。从而得出,10μg/mL的丝裂霉素C处理MEF 3 h,能够抑制细胞增殖,且不引起细胞死亡,能用来作ES细胞的饲养层,且可以冻存、复苏后直接用于培养胚胎干细胞。To discuss the inhibitory effect of mitomycin C on the proliferation of fibroblasts, the fibroblasts of Kunming mice were treated with 10 μg/mL mitomycin C after 1.5h, 2h, 2. 5h, 3h, 3.5h and 4h, respectively. The death of cells was observed by AO/PI stain and the proliferation of cells was monitored by MTT and PCNA stain. In addition, we compared the growth status of embryonic stem cells cultured on fi- broblast cell feeder layer which had been or not had been frozen-kept. The proliferation of fibroblasts could not be inhibited by treating with mitomycin C less than 3 hours. After 4 hours, a part of fibroblasts had lost their nucleoli and collapsed. There was no difference in the growth status of ESC cultured on fibroblast feeder layer which had been not had been frozen-kept. After 3 hours, by treating with 10 μg/mL mitomycin C, the fibroblast feeder layer cells were inhibited to grow but not to die. Therefore, they could be uesed as the feeder layer cells for the culture of ESC. Moreover, after frozen keping in liquid nitrogen and revitalised, they also could be used to culture ESC directly.

关 键 词：丝裂霉素C 成纤维细胞 细胞增殖 

分 类 号：Q813.1[生物学—生物工程] S852.1[农业科学—基础兽医学]