大鼠肝脏星形细胞的分离培养及鉴定

Isolation,culture and identification of hepatic stellate cells from normal rat liver

目的 建立大鼠肝星形细胞 (hepaticstellatecells,HSC)的体外分离及培养方法 ,为进行肝纤维化的实验研究提供技术支持。方法 选用SD大鼠经消化酶灌注肝脏后 ,用调整密度后的淋巴细胞分离液分离HSC。通过观察其自发荧光及显著其特征性结构的脂肪染色、细胞免疫化学染色等方法鉴定。结果 分离得到HSC ,细胞得率为 1～ 1 5× 10 7/只 ,存活率为98%。结论 成功分离及培养HSC ,且方法经济、简便。

Objective To study method of isolation and culture of rat hepatic stellate cell.Methods According to the method described in the references,a modified procedure for isolation and culture of rat hepatic stellate cells(HSC)was established with centrifugation on a single layer density cushion of Ficoll Hypaque(ρ=1 053).Results The yield of HSC using this method was about 1～1 5×10 7/per rat.Viability of the cells was more than 98%.All cells exhibited rapidly fading fluorescence under ultraviolet epi illumination.Desmin immunofluorescence and α smooth muscle actin(α SMA)staining as well as characteristic structure fat staining were all positive.Conclusion This work provides a good and economic method for culture of HSC which is very useful in the investigation of hepatic fibrosis.