改良小鼠肝星状细胞的分离、培养及性质鉴定

The improved method of isolation,culture and identification for mouse hepatic stellate cells

目的改进BALB/c小鼠肝星状细胞的分离、培养方法,提高其分离质量。方法采用胶原酶经下腔静脉手工逆灌注肝脏、optiprep密度梯度离心法分离小鼠肝星状细胞,含血清-DMEM培养星状细胞,台盼蓝拒染实验鉴定细胞活力;desmin、α-SMA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2×105个/鼠以上,纯度为93%～97%左右,肝星状细胞存活率为95%～97%。结论改良小鼠肝星状细胞的分离、培养方法,有利于原代小鼠肝星状细胞的生物学研究。

Objective To introduce a high efficient,convenient method for isolation and culture of mouse hepatic stellate cells（HSCs）.Methods The method for isolation of normal mouse HSC was established with collagenase in situ liver perfusion,and optiprep density gradient centrifugation.HSCs were cultured in DMEM medium with FBS.Desmin and α-SMA were examined for identification of the cells.Results The quantity of HSCs was above 2~105 cells per mouse.The purity and the viability of the cells were about 93~97 percent.Conclusion The result showed that this method of isolation for HSCs is more convenient and steadier;the relatively steady HSCs can be used in the biological study of primary culture of HSCs.