大鼠肝星状细胞的分离和培养

Isolation and culture of rat hepatic stellate cells

目的:本文参考Friedman等的不连续密度离心法并作改良,建立了一种经济、简便、可靠的分离大鼠肝星状细胞(HSC)的方法。方法:用链霉蛋白酶和胶原酶灌流大鼠肝脏,Nycodenz密度梯度离心分离HSC,并进行体外培养,结果:分离培养的HSC经台盼蓝拒染实验活细胞大于98％,细胞得率为3.95×10个/肝脏。结论:该方法简便、实用、稳定、可靠,而且本方法的建立为进一步研究HSC与肝纤维化的关系,尤其是细胞水平及分子生物学水平的研究奠定了基础。

Aim: HSC were isolated and purified according to the method of Friedman et al with some modifications, a modified simple and efficient density gradient centrifugation method was established in this paper. Methods:HSCs were isolated and cultured from liver of Wistar rats by in situ perfusion with pronase and collagenase and density gradient centrif ugation with Nycodenz. Results: HSCs were tested by Typan resistance tests,the living cells was about 98%,and the yield rate of HSC was about 3. 95@107 per rat. Conclusions: Our methods is convenients, practical, stable and reliable, and this modified method of preparing HSC may provide the basis for the study of liver fib rosis at the cellular and molecular level.