高原体外肝细胞分离方法的研究

目的:探索高原环境下动物肝细胞的体外分离方法。方法:采用胶原酶体外两步灌流法先进行肝叶灌流,再将所得肝组织悬液分别置37℃温育不同时间,对比观测肝细胞产量、存活率、纯度及细胞接种后生长情况的差异。结果:未经温育组及温育10min、30min、60min组肝组织悬液的肝细胞产量分别为0.85、2.67、3.84和4.29×105/g湿重肝组织;肝细胞存活率分别为97.6％、96.9％、88.3％及71.5％;肝细胞纯度分别为96.2％、95.4％、96.7％及 95.8％;前两组细胞贴壁生长和增殖情况较好,后两组则较差。结论:在高原环境下先采用肝叶体外两步灌流后冉行胶原酶短时间(10min左右)温育处理是一种既可获得较满意的细胞活性,同时又可显著提高细胞产量的分离肝细胞方法。

A METHOD TO COLLECT HEPATOCYTES AT HIGH ALTITUDE

Objective: To establish a method to collect hepatocytcs at high altitude. Method: The livers of rabbits were perfused with collagenase solution, and then the liver tissues and perfusion solution were incubated at 37C. at different times. The yield, viability and purity of hepatocytes and growth status of these cultured cells were observed. Result: The yields of hepatocytes colleted from the solution incubated for 0, 10, 30 and 60 min were 0.85, 2.67, 3.84 and 4.29 x 106 cells/g liver tissue, respectively. The viabilities were 97.6% , 96. 9%, 88. 3% and 71. 5%, and the purities were 96. 2%, 95. 4%, 96. 7% and 95. 8%, respectively. The growth status of the former two groups was better than that of the latters. Conclusion: Two- step eollagena.se liver perfu-sion and short incubation (about 10 min) is a method to collect hepalocyles from animal liver at high altitude, by which a large yield of hepatocytes as well as good viability can be obtained.