MSCs-肝细胞共培养上清对体外培养L02细胞的影响  

The effect of supernatant liquid form co-culture of MSCs with hepatocytes on L02 in vitro

作　　者：刘永[1,2] 胡文君[1] 蔡家利[2] 王英杰[1] 

机构地区：[1]第三军医大学西南医院感染病研究所,重庆400038 [2]重庆理工大学药学与生物工程学院

出　　处：《胃肠病学和肝病学杂志》2012年第3期253-256,共4页Chinese Journal of Gastroenterology and Hepatology

基　　金：国家自然科学基金资助项目(31070880)

摘　　要：目的以体外培养L02细胞为实验对象,了解骨髓间充质干细胞(MSCs)与肝细胞共培养上清的生物活性。方法采用两步酶分离法、全骨髓贴壁筛选法分别分离肝细胞、MSCs,按比例直接共培养,收集不同时段的共培养上清,用于人肝细胞系-L02细胞的培养。分别以单独培养的肝细胞上清、MSCs上清为对照,观察共培养上清对L02细胞活力和总蛋白合成的影响;同时观察其对L02细胞损伤模型的AST、LDH及细胞凋亡的影响。结果肝细胞上清、MSCs上清以及共培养上清对L02细胞的活力和增殖均有显著促进作用,表现为L02增殖活跃,总蛋白合成量增加;加入上述细胞上清的酒精损伤L02细胞AST、LDH释放和细胞凋亡显著低于未加细胞上清的对照组,其中共培养上清效果最为显著(P<0.05)。结论共培养上清对L02有明显的刺激增殖和损伤保护作用。Objective To investigate the biological activity of supernatant from co-culture of MSCs with hepatocytes. Methods In-situ collagenase two-step perfusion method and whole bone marrow adherent method were used to isolate hepatoctyes and MSCs of SD rats.Co-cultivation of hepatic cells and MSCs was done in vitro according to a certain ratio.The supernatant liquid form co-culture was collected to culture L02 cells,a human hepatic cell line,compared with the supernatant liquid of hepatic cells and MSCs which was cultured alone.Cell viability and total protein synthesis of L02 cells were investigated.The AST,LDH leakage and cell apoptosis of L02 cells damaged by 75% alcohol were also observed. Results Our results showed that the supernatant liquid of co-cultivation of hepatoctyes and MSCs,as well as the cultred hepatoctyes and MSCs could promote the cell viability and proliferation of L02 cells in vitro.Both the cell actively and the total protein synthesis of cultured L02 cells increased under the supernatant liquid.The amount of leakage of AST,LDH and cell apoptosis of damaged L02 cells by alcohol were lowered on the effect of culture supernatant liquid.Among the three supernatant liquids,the effect of the supernatant liquid of co-culture was most remarkable（P0.05）. Conclusion The supernatant liquid of co-culture can stimulate the proliferation of L02 cells and protect the cell injury significantly.

关 键 词：间充质干细胞 肝细胞 共培养上清 L02细胞 细胞活力 凋亡 

分 类 号：R575.2[医药卫生—消化系统]