小鼠胚胎成纤维细胞的分离培养

目的探讨小鼠胚胎成纤维细胞(MEF)的分离与培养。方法取BALB/c小鼠胚胎分离成纤维细胞,利用体外培养体系,对MEF的生长形态进行观察,并对传代细胞培养液、胚胎胎龄、胰蛋白酶浓度进行筛选。结果 MEF在体外为贴壁生长型细胞,第三、四、五代细胞纯度较高且增殖最为旺盛;添加血清的M199和DMEM均能较好的满足原代细胞的生长,两种培养液中细胞增殖的速度无明显差异;11.5～16.5d胎龄的小鼠胎儿MEF分离效果最好;0.1%胰蛋白酶消化MEF时间以8～11min为宜。结论通过对MEF分离培养影响因素的筛选,为MEF的进一步研究奠定了基础。

Study on Isolation and Culture of Mouse Embryonic Fibroblast

Objective To explore the isolation and culture of mouse embryonic fibroblasts （ MEF）. Methods The MEF were isolated from the embryos of BALB /c mouse using in vitro culture system,the growth morphology of the MEF were observed,and the passage cell culture medium,embryonic gestational age,the trypsin concentration were screened. Results The MEF in vitro was a kind of adherent cell,the third,fourth,fifth cell proliferation with high purity and the most exuberant,adding serum M199 and DMEM can satisfy the growth of primary cells, the speed of proliferation had no significant differences in two cell culture medium; the optimal gestational age for isolation of mouse embryonic fibroblasts was from 11. 5 to 16. 5 day. The optimal time for digesting MEF was 8 ～ 11 min with 0. 1% of trypsin. Conclusion By selecting the factors which influence of isolation and culture laid the foundation for further study of the MEF.