肌卫星细胞分离方法的改进及其应用

A modified method of isolation of satellite cell from murine skeletal muscle

目的:快速高效分离鼠类肌卫星细胞。方法:改进肌卫星细胞分离方法,即缩短分离时间并增加分离细胞数。结果:1.新分离法组织离体到细胞接种所需的时间为(1.9±0.47)h,与传统分离法(7.2±1.54)h 比较,细胞分离所需时间显著缩短(P<0.05)。2.新分离法每克组织分离获取的细胞数为(9.5±2.18)×10~6较传统分离法(4.8±1.81)×10~6显著增加(P<0.05),且细胞肌间线蛋白阳性率>97%。3.新分离法和传统分离法分离细胞在生长液中生长和分化液中分化成多核肌管的特性无显著差异(P>0.05)。结论:新分离法能快速高效分离获取纯度高的鼠肌卫星细胞。

Objective:To rapidly isolate satellite cells from murine skeletal muscle.Methods:The old method was modified to expedite the isolation process and increase of number of satellite cells from murine skeletal muscle. Results:(1)In the new isolation method,the cell isolation time was 1.9(0.47h which was markedly reduced com- pared to 7.2(1.54h for the old isolation method (P<0.05).(2)In the new isolation method,the number of isola- ted cell was 9.5(2.18(10~6/g.tissue which was markedly increased compared to 4.8(1.81 (10~6/g.tissue for the old isolation method (P<0.05).Furthermore,the Desmin+ rate was higher than 97%.(3)The characters of the isolated satellite cells by the new method were not different from that by the old method in regard to either pro- liferation or differentiation ability in both proliferation and differentiation media (P>0.05).Conclusion:The new method could be used to rapidly isolate and obtain more muscle satellite cells from routine skeletal muscle.