中文摘要：

目的：建立分离、纯化脐血间充质干细胞（HUMSCs）的方法，探讨肝衰竭患者血清对HUMSCs的诱导分化作用。方法：采用密度梯度离心和贴壁培养法相结合分离培养HUMSCs并鉴定，以舍重型肝病患者血清的低糖DMEM培养基诱导培养，不同时间点采用免疫组织化学方法（sP）检测甲胎蛋白（AFP）和白蛋白（Alb）的表达。结果：分离获得高纯度的HUMSCs，HUMSCs高表达CD44和CD29，不表达CD34，可以分化为脂肪细胞；SP结果表明，对照组HUMSCs不表达AFP和Alb，肝衰竭患者血清在培养7天时，大部分细胞呈AFP阳性表达（75．2±7．8）％，与对照组比较差异有显著性意义（P〈0．05）；诱导培养14天、21天和28天时AFP表达阳性率分别为（40．1±8．2）％、（10．4±3．5）％和（6．8±2．3）％；肝衰竭患者血清培养7天时仅少数细胞内可见Alb表达，表达阳性率为（9．8±2．7）％；培养14天、21天和28天，Alb表达阳性率分别为（25．4±5．2）％、（67．2±7．8）％和（83．4±7．9）％，与对照组比较差异有显著性意义（P〈0．05）。结论：密度梯度离心和贴壁培养法相结合是一种较好的分离纯化HUMSCs的方法；肝衰竭患者血清可以诱导HUMSCs表达AFP和Alb。

英文摘要：

Objective： To set up the method of isolate and purify human umbilical cord blood mesenchymal stem cells （HUMSCs） and investigate the inducing effect of serum of patients with severe liver disease. Methods： HUMSCs were isolated and purified by density gradient centrifugation and adherent culture. HUMSCs were induced in 20% liver disease patient serum in vitro, observed HUMSCs morphological change and differentiated character, detected albumin and AFP by SP. Results： High purity HUMSCs were isolated from umbilical cord blood. HUMSCs express CD44, CD29 and did not express CD34. HUMSCs might differenate to adipocyte cells in vitro. MSCs culture in L-DMEM medium with 20% serum of patients with severe liver disease , the cells become larger, showing epithelioid cell type. SP staining results： Alb and ck-18 was negative in the control group. Most cells were AFP positive after cultivated for 7 days （75.2 ± 7.8） %, compared with control group, there was significant difference （ P 〈 0. 05 ） ; AFP positive rate were respectively （ 40. 1 ± 8. 2 ） %, （ 10. 4± 3.5 ） % and （ 6. 8 ± 2. 3 ） % after cultivated for 14days, 21days and 28 days; Only a few cells were positive for7 days （9. 8 ± 2. 7） %, Alb positive rate were respectively （25.4 ±5.2）%, （67. 2± 7. 8）% and （83.4 ± 7.9）% after cultivated for 14d, 21d and 28 days, compared with control group and between different time points, there were significant difference （ P 〈 0.05 ） . Conclusion： A method for isolation and purification of HUMSCs in vitro has been established , Serum of patients with severe liver disease can induce the expression of AFP and Alb in HUMSCs.