小鼠c-Kit+lin-骨髓细胞移植生成肝细胞的实验研究

目的 评价小鼠c-Kit^＋Lin-骨髓细胞的肝干细胞潜能，为骨髓源性肝干细胞的临床运用提供可行性实验依据。方法 供体为纯系BALB／C雄性小鼠．c-Kit^＋Lin-骨髓细胞采用免疫磁珠分选法分离细胞。受体为行35Gy全肝照射处理后的同龄同系BALB／C雌性小鼠，立即移植供体c-Kit^＋lin-骨髓细胞。接受c-Kit^＋lin-骨髓细胞移植1月后，活杀取肝做常规病理学检查、Y染色体的原位杂交检查、甲胎蛋白与白蛋白的免疫组化检测。结果 移植1月后小鼠肝脏的组织结构已恢复正常．肝组织细胞中存在原位杂交和免疫组化均呈阳性反应的双阳性细胞。结论 小鼠c-Kit^＋Lin-骨髓细胞具有肝干细胞潜能，移植后可以在肝内定居并分化形成肝细胞。可做为肝病细胞移植疗法的种子细胞。

Differentiation of transplanted mouse c-Kit+lin- bone marrow cells into hepatocytes in vitro

Objective To ascertain whether mouse c-Kit Lin-bone marrow cells have the potential of hepatic stem cells.Methods c-Kit lin-bone marrow cells were isolated and purified by magnetic-activated cell sorting(MACS)from BALB/C male donor mice,and immediately transplanted into age-matched BALB/C syngeneic female mice with 35-Gy total liver irradiation.The recipients were sacrificed 1 month after the transplantation for pathological observation of the liver morphology.The presence of Y-chromosome was examined in the liver cells of the recipient by in situ hybridization(ISH),and alpha-fetoprotein(AFP)and albumin in the cells were detected by immunohistochemistry.Results The hepatocytes positive for Sry gene on Y-chromosome were identified 1 month after transplantation,and immunohistochemistry for AFP and albumin confirmed that the donor mice-derived cells were hepatocytes.Conclusion c-Kit lin-bone marrow cells have the potential of hepatic stem cells,which can reside and differentiate into hepatocytes in the liver after transplantaion.c-Kit lin-bone marrow cells can be used as the source cells of cell transplantation for liver disease.