【摘要】 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3×107/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。更多还原

【Abstract】 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 perfusion 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 3×107 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 subcultivation. Conclusion The rat HSCs are successfully isolated and cultured,suggesting our improvement is feasible.更多还原