日本血吸虫感染小鼠肝星状细胞的分离

目的 探索稳定、高效的血吸虫感染小鼠肝星状细胞的分离方法. 方法 采用Ⅳ型胶原酶体外灌注、11.5％ Optiprep密度梯度离心、CD11b磁珠阴性分选方法相结合分离血吸虫感染小鼠肝星状细胞. 结果 11.5％Optiprep密度梯度离心后细胞得率可达到(7.66±2.43)×106个/鼠,存活率为96％～98％.CD11b磁珠阴性分选后肝星状细胞得率可达到(3.9±1.33)×106个/鼠,CD11b、F4/80双阴性细胞纯度为90.8％～95.8％. 结论 本实验建立的血吸虫感染小鼠肝星状细胞分离方法稳定、高效,能够有效去除kupffer细胞的污染,为进一步研究HSC在血吸虫病肝纤维化中的功能提供基础.

Isolation of hepatic stellate cells from mice infected with Schistosoma japonicum

Objective To explore a steady and effective method for isolation of hepatic stellate ceils （HSCs）from mice infected with Schistosoma japonicum. Methods The method for isolating of infected mice HSC was established with Ⅳ collagenase in vitro liver recirculation perfusion,11.5% Optiprep density gradient centrifugation,and negatively sorted by CD11b magnetic microbeads. Results The quantity of HSC after density gradient centrifugation was （7.66 ± 2.43） ×10^6 cells per mice,the viability of the cells were 96% - 98%. After negatively sorted by CD11b magnetic microbeads the quantity of HSC was （3.9±1.33）×10^6 cells per mice,the purity of the CD11b, F4/80 - cells were 90.8%-95.8%. Conclusions The result showed that this method of isolating HSC of mice infected with Schistosoma japenicum was steady and effective, cultured HSC retain naive biological characteristics and can be used in future studies.