骨髓源性肝干细胞定向分化及脾内移植研究  

Differentiation induction and intrasplenical transplantation of bone marrow-derived liver stem cells

作　　者：蔡云峰[1] 陈积圣[1] 闵军[1] 魏菁[1] 何劲松[1] 莫隽全[1] 

机构地区：[1]中山大学孙逸仙纪念医院肝胆外科,广州510120

出　　处：《中华实验外科杂志》2004年第5期551-553,i001,共4页Chinese Journal of Experimental Surgery

摘　　要：目的　寻找骨髓源性肝干细胞的表面标记 ,进行定向分化及脾内移植研究。方法　流式细胞仪检测淤胆大鼠骨髓中干细胞群体的数量变化 ,寻找肝干细胞标记 ,免疫磁珠分离 ,进行体外诱导分化和肝再生模型的脾内移植 ,观察细胞形态的变化 ,免疫组织化学和免疫荧光技术检测白蛋白、AFP、CK8/18等肝细胞标记的表达。结果　淤胆鼠 β2微球蛋白阴性 (β2 m-)细胞数量较对照组数量明显增高 ,分别为 (6.17± 2 .70 ) %和 (0 .79± 0 .61) % (P <0 .0 1) ,β2 m-细胞体外培养及脾内移植均可出现肝细胞样细胞 ,白蛋白、AFP、CK8/18表达阳性。结论　β2m 细胞在体内外具有向肝细胞分化的能力 ,脾内移植是肝干细胞移植可供选择的部位之一。Objective To identify the surface marker of bone marrow-derived liver stem cells,isolate the stem cells,and investigate the differentiation and intrasplenical transplantation of the stem cells.Methods The variations of several stem cell surface markers,including β2-microglobulin negative (β 2m -),Thy-1 +,CD 34 +,Flt-3 +,IL-3R +,and c-kit + markers,were detected in the bone marrow of cholestatic rats by using flow cytometry.The significant cell populations (β 2m - cells) were then isolated using a magnetic bead cell-sorting procedure.The isolated cells were cultured in a system containing cholestatic serum and hepatocyte growth factor (HGF).They were also transplanted into the spleen of a rat model,which had been treated with retrorsine and 70% partial hepatectomy.The morphology of the cells was observed,and the expressions of albumin,AFP,and CK8/18 was detected by using immunohistochemistry and immunofluorescense.Results β 2m - cells were (0.79±0.61)% in the bone marrow of normal rats,significantly less than in the cholestatic rats [(6.17±2.70)%,P <0.01].After being co-cultured with cholestatic serum and HGF,the cells showed multilateral transformation and resembled hepatcytes morphologically.In the spleen of the model rats,hepatocyte-like cells appeared.Both the cultured cells and the transplanted cells expressed albumin,AFP,and CK8/18,all known to be the characteristic markers of hepatocyte.Conclusion β 2m - cells have the ability to trans-differentiated into hepatocyte both in vivo and in vitro.Spleen can be a suitable organ for the transplantation of bone marrow-derived liver stem cells.

关 键 词：骨髓源性肝干细胞 定向分化 脾内移植 细胞形态 免疫组织化学 免疫荧光技术 

分 类 号：R657.3[医药卫生—外科学]