一种改进的分离纯化人肝脏星形细胞的方法

A modified method for isolation and purification of human hepatic stellate cells

目的 改进传统的分离纯化人肝脏星形细胞 (HSCs)的方法。方法 在传统的分离人HSCs方法的基础上 ,省略链霉蛋白酶消化 ,仅用Ⅳ型胶原酶消化 ,采用低速离心去除肝脏实质细胞 ,将获得的肝脏非实质细胞进行两次换液及传 1代培养 ,观察培养过程中细胞形态及蛋白标志物的改变。结果 经传 1代培养 2 4h后 ,所分离得到的人HSCs纯度接近 10 0 %。结论 所改进的分离人HSCs的方法具有操作简便 。

Objective To improve the method for isolation and purification of human hepatic stellate cells (HSCs). Methods A commonly used method for isolation of human HSCs was modified. The new method only involved type Ⅳ collagenase perfusion of the liver in which hepatic cells were removed by low speed centrifugation and the hepatic nonparenchymal cells were plated and the culture medium was changed twice within 24 h. The cells were subcultured in primary culture in 7-10 d. The morphology and protein marker of the cells were observed at different intervals in the cultures with phase contrast microscopy and immunocytochemical staining. Results The purity of the isolated HSCs was almost 100% in 24 h after first subculture. Conclusion The modified method for the isolation of human HSCs is simple and effective.