改良的大鼠肝星状细胞分离方法

An improved method for isolation of hepatic stellate cells

目的:探讨一种简便、高产的大鼠肝星状细胞(HSC)分离方法,为研究肝细胞相关实验提供细胞原材料。方法:应用悬浮缓冲液校正Nycodenz密度梯度分离液,使其浓度为11.3%(w/v),采用单层一步梯度离心法分离HSC。采用台盼蓝染色排斥法鉴定细胞活率,desmin免疫细胞化学染色法鉴定HSCs纯度。结果:肝星状细胞得率每只大鼠约(4.0±0.5)×107个,细胞活力达到90%,细胞纯度达到95%。结论:本法简便、高产,是一种理想的分离肝星状细胞的方法,值得推广。

Objective： To investigate a simple and efficient method for isolation of hepatic stellate cells （HSC）, this may provide raw material for studying the liver fibrosis at the cellular level. Methods： Cell suspension and Nycodenz cell separating medium were mixed, and the concentration of the fluid was adjusted to be 11.3% （w/v）, HSC were isolated by momolayer gradient centrifugation method. The cells viability was identified by Trypan blue exclusion staining. The purity of HSCs was identified by the expression of desmin immunocytochemistry. Results： The yield rate was about （4.0±0.5）×10^7 per rat, the survival and the purity rate of HSC were 90%, 95% respectively. Conclusion： The simple and efficient method is an ideal one for the isolation and culture of rat HSCs, which is worth spreading.