一种简单分离、传代培养成年小鼠肝细胞的方法

A Simplified Methid in Isolation and Culiuring adult Mouse liver cells

目的 寻找肝细胞分离、传代培养更简单方便的方法。方法 应用不同胰酶浓度、不同消化时间消化成年小鼠肝组织后进行组织块培养 ,观察细胞的贴壁率、产量及活力。结果 胰蛋白酶分离成年小鼠肝细胞 ,于 MEM(含 2 0 %小牛血清 )培养液中 ,37℃密闭培养 ,生长情况良好 ,12小时左右即可完全贴壁 ,并已传至第二代。胰酶浓度为 0 .15 % ,消化时间15 m in,细胞产量及活率较高 ,每 10 g体重细胞产量为 0 .84± 0 .0 8× 10 7,细胞活力为 (90 .6 6± 5 .71) % ,贴壁生长 48小时细胞倍增 PD值为 0 .86± 0 .0 5。结论 胰蛋白酶消化、组织块贴壁法是成年小鼠肝细胞分离、培养传代的简便方法 ,不需要特殊装置 ,酶耗费少 。

Objective To establish a more simplify and convenience method for isolate and culture liver cells. Methods Using a various concentrations of trypsin and incubation times to digest liver tissue,and then proceed tissue culture The cells anchoring rate,output and vital force were observed Results Liver tissue from RM mouse isolated by trysin digestion grow well in MEM with 20%fetal bovine After 12hours,the liver cells have been subcultured to second generations,the parenchymal cells nestle closely to the bottle wall The hepatocytes in the tissue digested for 15 minutes by 0 15% trypsin guew the best ,and aoutput of 0 84±0 08*10 7cels/10g was obained,liver cels vital force were 90±5 75% During 48 hours the cells population dubling(PD)was 0 86±0 05 Conclusion Digested by trypsin,tissue anchoring culture was a simplify and conveience mehtod for isolate and culture hepatocyte It doesn't need special device,enzyme consume was very few,the cell output may meet the need of common experiment