摘要
目的改良分离培养大鼠肝星状细胞的简便方法.方法经过门静脉插管、推注肝素稀释液、肝脏灌流、胶原酶离体循环消化、过滤、离心分离等步骤,分离肝星状细胞,省去了链霉蛋白酶E消化过程,采用台盼蓝染色排斥法鉴定细胞活率,des血n免疫细胞化学染色法鉴定HSCs纯度.结果每只鼠肝HSCs的得率为0.5～1.0×107,纯度为(92.2±2.0)%,活率为(97.8±0.3)%,数量及活率均符合实验要求.结论该方法简便、廉价、实用,值得进一步探索.
To improve a convenient method of isolating and culturing rat hepatic stellate cells (HSCs). Rat hepatic stellate cells (HSCs) were isolated by the following steps: intubating portal vein, injecting diluted heparin, liver perfusion, collagenase liver circulating perfusion in vitro, cells filtering, density gradient centrifugation. But the course of liver cells digestion by pronase E was left out. The cells viability was identified by Trypan blue exclusion staining. The purity of HSCs was identified by the expression of desmin immunocytochemistry method. The yield rate of HSCs was 0.5-1.0×107 per rat . The purity was (92.2±2.0)%.The cells viability was (97.8±0.3)%. [Conclusions] The modified method of isolating and culturing HSC is simple,cheap , reliable and worth further research.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2005年第2期205-207,共3页
China Journal of Modern Medicine
关键词
大鼠
星状细胞
细胞分离
细胞培养
rat
hepatic stellate cells
cells isolation
cells culture
