肝纤维化小鼠原代肝细胞的分离培养

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中图分类号：R575.2 文献标志码：A DOI ： 10.3969/j . issn.1003-1383.2025.12.003

【Abstract】ObjectiveTo establishareliable method for isolatingandpurifying primary liver celsfrom mice with high vitalityand purityofliverfibrosis.MethodsTheclasscal two-stepcollgenase perfusion method wasimproved byusing inferior vena cava perfusion，intermittent clamping of the portal vein perfusion，and 50% Percoll density-gradient centrifugation forpurificationof hepatocytesuspension.Thepurified hepatocyteswereculturedin vitro，andimmunofluorescentstainingof Albuminand HNFlαwasperformed.ResultsThesurvivalrate of primary livercelsisolatedandpurified from liver fibrosis mice could stably reach （92.93±1.93）% ，with an average total number of live cells of （1-2）×10′ per adult mouse.Purified cellsadhered to the substrate and began to spread within 2h of in vitro culture. Conclusion A stable technique forisolating and culturing primary livercelsfrom micewith high purityandvitalityin liver fibrosisissuccesfully established.

【Keywords】liver fibrosis；mouse hepatocytes； high purity hepatocytes； in vitro culture technique

肝纤维化是肝脏在长期或反复损伤刺激下启动的一种复杂的修复性反应，其特征性病理改变为细胞外基质成分，特别是I型和Ⅲ型胶原蛋白的异常积聚和重构，是正常肝组织恶变过程中，进展为肝脏肿瘤的重要转变阶段[1]。（剩余7898字）