肝细胞生长因子对人脐带间充质干细胞体外增殖的影响  

Effect of Hepatocyte Growth Factor on Proliferation of Human Mesenchymal Stem Cells in Vitro

作　　者：王慧 刘佳星 周燕[1] 谭文松[1] WANG Hui;LIU Jiaxing;ZHOU Yan;TAN Wensong(State Key Laboratory of Bioreactor Engineering,East China University of Science and Technology,Shanghai 200237,China)

机构地区：[1]华东理工大学生物反应器工程国家重点实验室,上海200237

出　　处：《华东理工大学学报（自然科学版）》2022年第6期790-796,共7页Journal of East China University of Science and Technology

基　　金：国家重点研发计划项目“生物医用材料研发与组织器官修复替代”(2018YFC1105800)。

摘　　要：为了改善无血清培养基促细胞增殖能力,探讨了肝细胞生长因子(Hepatocyte growth factor, HGF)对间充质干细胞(Mesenchymal stem cells, MSCs)体外增殖的影响。采用人脐带来源的MSCs,在实验室前期自制的无血清培养基SFM-O中添加不同浓度的HGF,采用CCK-8法测定细胞增殖;使用流式细胞术检测细胞周期,通过Western blot法检测细胞周期蛋白Cyclin D1及相关信号通路蛋白的表达。结果表明:在自制的无血清培养基中添加10 ng/mL的HGF能够有效促进MSCs的增殖;HGF能增强MSCs的整合素表达,进一步通过激活FAKAKT-mTOR信号通路促进Cyclin D1蛋白合成,加速细胞周期从G1期到S期的转变,从而促进MSCs增殖。Mesenchymal stem cells(MSCs) are promising candidates for cell therapy and tissue engineering.However, the currently self-developed serum-free medium(SFM) has problems in obtaining a sufficient number of clinically available cells in vitro. A breakthrough may be found by fully studying the key medium components which influence cell behavior. In this study, the effect and mechanism of hepatocyte growth factor(HGF) on cell proliferation were investigated. The self-made serum-free medium at the early stage from laboratory was used as the control group,adding different mass concentrations of HGF and finding the most effective one. Cell counting kit-8 and growth curve were used to evaluate its proliferation. And cell cycle transition was also analyzed by flow cytometer. Moreover,western blotting was used to detect the expression levels of cyclin D1, integrin and the related signaling pathway. The results showed that 10 ng/mL HGF could significantly increase MSCs proliferation and viability in SFM. The effect of HGF occurred via active FAK-AKT-mTOR pathway and increased the synthesis of cyclin D1 protein, accelerating cell cycle from G1 phase to S phase progression, and promoting the proliferation of MSCs. HGF could enhance the expression of integrin, which is helpful to signaling transduction. After adding the inhibitors of FAK and AKT,phosphorylation of the corresponding protein decreased. Finally, after adding HGF, MSCs could still express surface marker including CD44, CD73, CD90, and CD105, and led to osteogenesis and adipogenesis differentiation. Thus,these findings have important implications for utilizing HGF to regulate cell behavior and expand clinical-grade MSCs in vitro.

关 键 词：间充质干细胞 肝细胞生长因子 无血清培养基 增殖 AKT 

分 类 号：Q291[生物学—细胞生物学]