摘要
目的 确定骨髓源性肝干细胞的表面标记 ,体外培养诱导其分化为肝细胞。方法 采用流式细胞仪检测多种大鼠肝损伤模型骨髓中干细胞标记的细胞群体的数量变化 ,寻找可能的肝干细胞 ,免疫磁珠分离各骨髓干细胞标记的细胞群体 ,进行体外培养诱导分化 ,观察细胞形态的变化 ,免疫组化技术检测清蛋白 (白蛋白 ) ,AFP ,CK 8/ 18等肝细胞标记的表达。结果 各模型大鼠骨髓内 β2微球蛋白阴性 (β2 m- )细胞显著增高 ,体外培养经诱导后呈多角形细胞表现 ,清蛋白 ,AFP ,CK 8/ 18表达阳性。其他干细胞类型细胞数量变化小 ,体外培养未见肝细胞样变化。结论 β2 m- 细胞的数量随肝损伤而变化 ,在体外具有向肝细胞分化的能力 。
Objective To identify the surface marker of bone marrow-derived liver stem cells and to isolate the stem cells, to investigate the differentiation of the stem cells. Methods The quantitative variations of the cells with stem cell surface markers, including β 2-microglobulin negative (β 2m -), Thy-1 +,CD34 +,Flt-3 +,IL-3R +,and c-kit + markers, were detected by using flow cytometry in the bone marrow of several rat models with liver injury. Each stem cell population was 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). The morphology of the cells was observed, and the expressions of albumin, AFP, and CK8/18 were detected with immunohistochemistry technique. Results β 2m - cells elevated significantly in each of the rat models.After being co-cultured with cholestatic serum and HGF, β 2m - cells showed multilateral transformation and resembled hepatcytes morphologically. The differentiated cells expressed albumin, AFP, and CK8/18, all known to be the characteristic markers of hepatocyte. The other cell population showed little quantitative changes, and did not express the same proteins. Conclusions The quantitative variation of β 2m - cells corresponds to the severity of liver injury. β 2m - cells have the ability to trans-differentiated into hepatocytes in vitro. They might be the marker of liver stem cells.
出处
《中国普通外科杂志》
CAS
CSCD
2003年第4期287-290,共4页
China Journal of General Surgery
