改良大鼠肝星状细胞分离及性质鉴定

The improved method of isolating and identifying rat hepatic stellate cells

目的探索经济、稳定的肝星状细胞提取和培养方法。方法采用胶原酶改良原位循环灌流、18%Nycodenz密度梯度离心法分离大鼠肝星状细胞,胎牛血清DMEM培养星状细胞;desmin、αSMA和ICAM1免疫细胞化学检测进行星状细胞性质鉴定。结果肝星状细胞的得率稳定在3.5×107/肝以上,纯度为96%～98%左右,存活率为96%～98%。结论建立了经济、稳定的大鼠肝星状细胞提取和培养方法;传代培养可获得较为稳定的细胞系。

Objective To investigate more economical and steadier method for the isolating and culturing of hepatic stellate cells (HSCs). Methods The method for isolating of normal rat HSC was established with collagenase in situ liver recirculation perfusion, and 18% Nyco denz density gradient centrifugation. HSCs were trained in digestion DMEM culture medium. De smin, dt-SMA and ICAM-1 were examined for identifing the cells. Results The quantity of HSC was above 3.5～10~7 cells per rat. The purity and the viability of the cells were about 96～98 percent. Conclusion The result showed that this method of isolating HSC was more economical and steadier; the relatively stead y cell line was acquired by subculturing.