[小鼠胚胎成纤维细胞的分离与培养]
[Isolation and culture of mouse embryonic fibroblast].
作者信息
Zhang Yi, Zhao Liansan, Wang Chengxiao, Lei Bingjun
机构信息
Infectious Diseases Department, West China Hospital, Sichuan University, Chengdu 610041, China.
出版信息
Sichuan Da Xue Xue Bao Yi Xue Ban. 2003 Apr;34(2):344-6.
OBJECTIVE
To establish a stable culture system of mouse embryonic fibroblast(MEF).
METHODS
We isolated MEF from the embryos of BALB/c mouse and investigated the morphology, growth curve and cell cycle of MEF in vitro. The effects of the gestational age (days) of mouse embryos and the time of digestion on the isolation and culture of MEF were assessed.
RESULTS
The mouse embryo of 13.5 d is better than that of 10.5 d and 18.5 d on the isolation of MEF. MEF in vitro is a kind of adherent cell with good ability for proliferation in passage 3. In room temperature, the digestive time of 0.25% trypsin should be 3-5 min.
CONCLUSION
MEFs(P3) are quite suited to be a feeder layer of embryonic stem cells.
目的
建立稳定的小鼠胚胎成纤维细胞（MEF）培养体系。
方法
从BALB/c小鼠胚胎中分离MEF，体外研究MEF的形态、生长曲线和细胞周期。评估小鼠胚胎的胎龄（天）和消化时间对MEF分离和培养的影响。
结果
在MEF分离方面，13.5天的小鼠胚胎优于10.5天和18.5天的胚胎。体外培养的MEF是一种贴壁细胞，传代3次时增殖能力良好。在室温下，0.25%胰蛋白酶的消化时间应为3 - 5分钟。
结论
MEFs（P3）非常适合作为胚胎干细胞的饲养层。
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