人脐带源间充质干细胞静脉移植治疗大鼠肝纤维化

背景：近来国内外研究证明，来自其他组织的干细胞能够归巢到肝脏，并可能参与肝组织的再生，这为发展干细胞治疗肝脏疾病提供了新的希望。目的：探究人脐带源间充质干细胞的分离、培养方法，观察脐带间充质干细胞移植对大鼠肝纤维化模型的修复作用，为脐带间充质干细胞的临床应用提供可靠的理论依据。方法：自然贴壁法分离、纯化人脐带间充质干细胞并进行体外培养和扩增，用皮下多点注射CCl₄制备肝纤维化大鼠模型。将22只模型大鼠随机分为模型损伤组(n=11)和细胞移植组(n=11)。细胞移植组在模型制备成功后的第1，2，3周经尾静脉给予1×10⁶脐带间充质干细胞治疗，4周后将大鼠处死，收集各组大鼠血液检测肝功能；摘取肝脏行苏木精-伊红染色，观察病理变化；免疫组化法观察库普弗细胞的数量及分布；免疫组化法观察治疗组脐带间充质干细胞的定位情况。结果与结论：脐带间充质干细胞经尾静脉移植入肝硬化大鼠后，大鼠的肝功能均明显改善，与对照组比较，差异有显著性意义(P < 0.05)；肝组织苏木精-伊红染色提示，肝纤维化程度明显改善；免疫组化法观察库普弗细胞的数量提示，库普弗细胞数量明显减少；免疫组化方法利用抗BrdU抗体在治疗组大鼠肝脏观察到BrdU标记的脐带间充质干细胞。说明脐带间充质干细胞移植可以改善大鼠外周血液的血生化特性和肝的组织学结构。中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程全文链接：

Vein transplantation of human umbilical cord-derived mesenchymal stem cells for treatment of hepatic fibrosis in rats

BACKGROUND:Recent studies verified that other tissues-derived stem cells can be homing to the liver, possibly participate in the regeneration of liver tissues, which provides new hope for stem cells in treatment of liver disease. OBJECTIVE:To observe isolation and culture of human umbilical cord-derived mesenchymal stem cells, to observe the repairing effect of umbilical cord-derived mesenchymal stem cell transplantation on hepatic fibrosis, and then to provide a reliable theoretical basis for further clinical application of umbilical cord-derived mesenchymal stem cells. METHODS:Human umbilical cord-derived mesenchymal stem cells were isolated and purified by natural adherent method, and then cultured and amplified in vitro. A rat model of hepatic fibrosis was prepared using subcutaneous multi-point injection of CCl₄. A total of 22 rat models were randomly assigned to model injury group (n=11) and cell transplantation group (n=11). At 1, 2 and 3 weeks after model induction, the rats in the cell transplantation group were treated with 1×10⁶ umbilical cord-derived mesenchymal stem cells via caudal vein. Four weeks later, the rats were sacrificed. Blood of rats was collected from each group to detect liver functions. The liver was removed to receive hematoxylin-eosin staining, and pathological changes were observed. The number and distribution of Kupffer’s cells were observed using immunohistochemistry. The localization of umbilical cord-derived mesenchymal stem cells from treatment group was observed using immunohistochemistry. RESULTS AND CONCLUSION:After umbilical cord-derived mesenchymal stem cells were infused into rats with  cirrhosis via caudal vein, liver function was significantly improved, which showed significant differences as compared with the control group (P < 0.05). Hematoxylin-eosin staining revealed that hepatic fibrosis was apparently improved. Immunohistochemistry results demonstrated that the number of Kupffer’s cells was obviously reduced, and BrdU-labeled umbilical cord-derived mesenchymal stem cells were visible in rat liver of the treatment group using anti-BrdU antibody. These results suggested that umbilical cord-derived mesenchymal stem cell transplantation could improve biochemical characteristics of rat peripheral blood and histological structure of liver.