SD大鼠肝脏星状细胞分离培养及方法的改进

目的：建立高效、简便的肝星状细胞分离方法.方法：采用改良原位灌流、12%Optiprep密度梯度离心法分离大鼠肝星状细胞,desmin和α-SMA 免疫细胞化学检测鉴定星状细胞性质.结果：肝星状细胞的得率稳定在3×10^7/肝以上,纯度为95%左右,存活率为98%,传代培养10代以上,细胞增殖及转分化状态仍良好.结论：肝星状细胞分离培养成功,对分离方法的改进措施可行.

Improvement in isolation and culture of SD rat heptic stellate cells

Objective To introduce a high efficient, convenient method for isolation and culture of rat hepatic stellate cells （HSCs）. Methods Rat HSCs were isolated by modified in situ peffusion with pronase and collagenase followed by density gradient centrifugation with 12% Optiprep. The purity of HSCs was identified by the expressions of desmin and α-SMA using immunocytochemistry method. Results The harvest rate of hepatic stellate cells was 3×10^7 cells per liver with the purity of 95% and viability of 98%. Cell proliferation and transdifferentiation remained in good condition after 10 generations of serial subcuhivation. Conclusion The rat HSCs are successfully isolated and cultured, suggesting our improvement is feasible.