ICR小鼠胚胎成纤维细胞的培养及饲养层的制作

目的：建立ICR小鼠胚胎成纤维细胞饲养层培养体系，用于胚胎干细胞建系及研究。方法：选取不同胎龄的鼠胚原代细胞分离成纤维细胞，制作饲养层，使用不同浓度丝裂霉素C处理，以细胞形态、生长曲线、囊胚种植情况为饲养层评价指标。结果：ICR小鼠胚胎成纤维细胞可在胎龄12．5～14．5d进行原代细胞分离；12．5d为最佳分离时间；13．5和14．5d分离的细胞需在3代以后使用；6代以前细胞可用于饲养层制作；20mg/L丝裂霉素C作用2．5h可达到较好的处理效果。结论：ICR小鼠可用来分离、培养胚胎成纤维细胞，用于胚胎干细胞饲养层的制作。

Culture of ICR mouse embryonic fibroblasts and production of feeder layers for embryonic stem cells

Objective: To establish feeder layer culture system of ICR mouse embryonic fibroblast(MEF) cells for isolation and culturing of embryonic stem(ES) cells. Methods: ICR mouse fetuses of different gestational ages were chosen to isolate mouse MEF as the feeder layers of ES cells.Different concentrations of mitomycin C were used to pretreat feeder layers.The effects of feeder layers were assessed by cell morphology,growth curves and the development of blastocysts. Results: MEF cells could be isolated from 12.5-14.5 d pregnant mice;MEF cells from 12.5 d pregnant mice grew better than those from 13.5 d or 14.5 d ones.Cells before 6th passage could be used to product feeder layers.The optimal concentration of mytomycin C to inhibit proliferation of MEF was 20 mg/L for 2.5 h. Conclusion: ICR mice can be used for isolation of MEF to product Feeder Layers for ES cells.