• 实验研究 •    

间充质干细胞异种移植治疗肝损伤的实验研究

王福财,刘玉琳,傅颖媛,施桥发   

  1. 南昌大学医学院
  • 收稿日期:2009-12-18 修回日期:2010-02-24 出版日期:2010-06-15 发布日期:2010-06-15
  • 通讯作者: 傅颖媛

The Study of Mesenchymal Stem Cells Heterogenic Transplanted to Improve Liver Damage

  • Received:2009-12-18 Revised:2010-02-24 Published:2010-06-15 Online:2010-06-15

摘要: 目的 比较研究小鼠骨髓间充质干细胞(MSCs)和单个核细胞(MNCs)在四氯化碳(CCl4)诱导性大鼠肝损伤治疗中的作用,为临床进行骨髓干细胞异种移植治疗肝脏疾病提供新策略。方法 无菌条件下取雄性BALB/c小鼠骨髓分离MNCs,培养纯化及流式细胞仪检测MSCs;采用CCl4制作雌性Wistar大鼠肝损伤模型,肝损伤大鼠随机分为MSCs移植组、MNCs移植组和肝损伤对照组。两细胞移植组在CCl4损伤后立即经尾静脉注入小鼠MSCs或MNCs,6周后处死小鼠,检测大鼠肝损伤程度和小鼠MSCs或MNCs植入情况。结果 流式细胞仪结果显示:小鼠MSCs CD29、CD44和CD90表达阳性,而CD34和CD45表达阴性;肝功能检测和肝组织学观察显示:MSCs移植组肝损伤修复程度明显好于MNCs移植组和肝损伤对照组;PCR方法证实只有MSCs移植组肝内有小鼠Y染色体阳性细胞存在。结论 体外分离培养及扩增的小鼠MSCs移植可改善小鼠CCl4导致的肝损伤。

关键词: 间充质干细胞, 单个核细胞, 异种移植, 肝损伤

Abstract: Objective To comparative study the effect of mesenchymal stem cells (MSCs) and mononuclearcells(MNCs) from mice bone marrow in liver damage of carbon tetrachloride (CCl4)-treated rat, and to provide a new method for bone marrow stem cells transplantation to care liver disease. Methods Bone marrow was collected from male BALB/c mice under sterile condition. MNCs were isolated, and MSCs were cultured, purified and identified by using flow cytometry. Female Wistar rat were challenged with CCl4 to make liver damage models. The rat with liver damage were randomly divided into MSCs transplanted group, MNCs transplanted group and liver damage control group. MSCs or MNCs were infused immediately through the tail veins after CCl4 treated in these cells transplanted mice. After six weeks, we assessed liver damage and mice cell engraftment. Results According to flow cytometry assay, MSCs were positive for CD29, CD44 and CD90, while negative for CD34 and CD45; Liver function and Histological examination suggested that liver damage recovery was much better only in these MSCs transplanted mice; PCR analysis revealed that mice Y-chromosome-positive cells only can be founded in these MSCs transplanted rat. Conclusions Mice MSCs can be isolated, cultured and expanded in vitro. Transplanted to CCl4-treated rat, they can improve liver damage.

Key words: mesenchymal stem cells, mononuclearcells, heterogenic transplantation, liver damage