摘要
目的 :介绍一种合理、实用、便捷的肝星状细胞 (HSCs)原代培养方法。方法 :对SD大鼠进行肝脏原位灌注、酶消化 ,并进行间质细胞密度梯度离心。使用 2 0 %、35 %和 5 0 %三层Percoll密度梯度分离纯化HSCs,分析其纯度、活力 ,并根据原代和传代HSCs的特征进行鉴定。结果 :每只鼠肝HSCs原代培养得率为(1.5～ 2 )× 10 7个细胞 ,纯度为 (97± 2 ) % ,细胞活力为 (98± 0 .6 ) %。原代和传代HSCs都表达Desmin ,只有传代 7d的HSCs表达α SMA。电镜见细胞浆内大量脂滴。结论 :本方法简单、实用、方便 ,所得结果优于其他方法。
Objective:To introduce an appropriate,useful and convenient method for HSCs primary culture in vitro.Methods:Using the methods as usual,in situ liver perfusion,collagenase Ⅳ and pronase E digestion and density gradient centrifugation,primary non-parenchymal cells were separated by 20%,35%,50% percoll in diverse concentrations in SD male rats. The total yield,purity and activity of HSCs were analyzed. Also some characteristics of primary and passaged cells were detected.Results:HSCs yield was(1.5～2)×10 7 per rat liver,purity and activity of cells were(97±2)%,(98±0.6)% respectively. Immunochemical detections showed both primary and passaged HSCs were Desmin positive in microscope analysis,but only passaged HSCs presented α-SMA positive. A large amount of lipid like droplets could be seen in the cytoplasm under the TEM.Conclusion:The method in this study is more simple,useful and convenient comparing with other methods introduced by some references,it guarantees the purity,activity of HSCs. Using this method can save ample money and time in preparing primary HSCs in vitro.
出处
《肝胆胰外科杂志》
CAS
2003年第4期226-229,共4页
Journal of Hepatopancreatobiliary Surgery
基金
上海市科委课题资助项目 ( 0 2 410 70 10)
关键词
原代培养
肝星状细胞
大鼠
primary culture
hepatic stellate cells
rat
