成肌细胞体外培养中两种纯化方法的比较

The Comparison of Two Purification in Myoblasts Culture

目的:比较成肌细胞体外培养中两种纯化方法的优劣。方法:将鸡胚成肌细胞进行体外培养,分别用差速贴壁法和Percoll液梯度离心法纯化成肌细胞。每组培养皿中放置3块盖玻片。每天观察两组细胞的增殖情况,细胞贴壁后第7d更换融合培养基,继续培养6d,取出盖玻片进行HE染色,观察细胞融合情况。结果:两组细胞都有融合,但梯度离心法纯化后培养的细胞融合率较贴壁组高,差异有统计学意义(P<0.01)。结论:梯度离心法纯化的成肌细胞培养后细胞纯度更高,建议用此法进行成肌细胞的纯化。

Objective：To compare the advantage and disadvantage of two different purification in myoblasts culture. Methods： Myoblasts of chick embryo were cultured in vitro. Purifying the myoblasts by differential adhesion or Percoll density gradient centrifugation. Laying three cover slips in each culture dish and observing proliferation behavior of myoblasts every day. The syncretic medium was added in culture dish in seven days after adhesion of myoblasts and then culture was kept on for 6 days. Cover slips were taken out and HE staining was operated. Observing the situation of cell fusion. Results：The cell fusion exists in two groups,but the ratio of cell fusion in density gradient centrifugation group is higher than it in the other group, and it shows a obviously difference between two groups（P〈0.01）. Conclusion：The cell purity which obtained by the method of density gradient centrifugation is higher and we advise to use this way to sublime the myoblasts.