东方田鼠胚胎成纤维细胞的分离培养及生物学特性

目的对影响东方田鼠胚胎成纤维细胞（MfEF）分离和培养的因素进行探索,并观察其生物学特性。方法取室内繁殖饲养不同胎龄的东方田鼠胚胎分离成纤维细胞,通过原代和继代培养,分析比较不同胎龄、不同血清浓度、不同胰蛋白酶浓度等因素对MfEF分离及培养的影响,观察MfEF的生长形态及其生物学特性。结果 MfEF为贴壁型生长,细胞形态多样,呈梭形、不规则多边形;采用0.125%的胰蛋白酶室温消化12～13 d胚胎组织5 min,以DMEM培养基添加15%小牛血清分离培养MfEF的效果最佳;MfEF2～7代增殖最旺盛。结论获得了实验室分离、培养MfEF的有效方法 ,为进一步深入研究东方田鼠抗日本血吸虫机制以及开展不同动物成纤维细胞间比较研究奠定了基础。

Isolation, Culture and Biological Characteristics of Embryonic Fibroblasts from Microtus fortis

Objective The aim of this study was to isolate,culture,and characterize embryonic fibroblasts from Microtus fortis（MfEF）.Methods The primary embryonic fibroblasts were isolated from Microtus fortis fetus at different gestational ages.According to the influence of different serum and trypsin concentrations and different gestational ages,the growth behavior and the biological characteristics of Microtus fortis embryonic fibroblasts（MfEF） were studied.Results The MfEF in vitro appeared as adherent cells with good ability for proliferation in passages 2-7.The optimal gestational age for isolation of MfEF was from day 12 to 13.At the temperature of 20℃ to 25℃,the optimal time for digestion of fetal tissue to obtain fibroblasts was 5 min with a digestion solution containing 0.125% of trypsin.The MfEF were cultured in DMEM（low glucose） supplemented with 15% FBS.Conclusion Embryonic fibroblasts of Microtus fortis have been successfully isolated and cultured,which lay the foundation for further studies on Microtus fortis against Schistosma japonicum infection and for comparative study of fibroblasts between different animals.