关键词: 小鼠, 胚胎成纤维细胞, 分离, 培养

Abstract: 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.

Key words: Mouse, Embryonic fibroblast, Isolation, Culture