大鼠脂肪间充质干细胞的鉴定及肝内移植的研究

目的:从脂肪组织中获取间充质干细胞(ADMSCs)并验证其多向分化潜能,探讨ADMSCs在肝再生中的作用。方法:获取大鼠脂肪组织,用胶原酶消化法获取干细胞,并进行体外扩增、传代,取第3代细胞分别用不同诱导培养液进行成骨、成脂诱导,诱导后通过细胞形态学和特殊染色观察诱导效果。用PKH26标记细胞,制作部分肝切除模型,将标记的自体ADMSCs经门静脉植入体内,2周后切下取肝脏制成冰冻切片,荧光显微镜观察植入细胞在肝脏的定位,免疫荧光染色观察其白蛋白的表达。结果:从脂肪组织中分离出的细胞能在体外大量扩增,能被诱导分化为成骨细胞、脂肪细胞,ADMSCs移植2周后,可见PKH26标记细胞散在分布于肝内,免疫荧光染色显示标记细胞白蛋白染色阳性。结论:大鼠脂肪组织中可以获取具有多向分化潜能的间充质干细胞,该细胞在肝再生环境中能向肝细胞分化,参与肝再生。

Identification and Intrahepatic Transplantation of Adipose-Derived Mesenchymal Stem Cells

Objective:To verify whether the cells derived from rat adipose tissue are mesenchymal stem cells which have the mul- ti-directional differentiation ability and to investigate whether adipose tissue-derived mesenchymal stem cells(ADMSCs)participate in liver regeneration after partial hepatectomy.Methods:Rat adipose tissue was harvested,stem cells were isolated using the method of col- lagenase enzyme digestion,then the cells were amplified,subcultivated,and the third passage cells were induced into adipose cells,os- teoblast and neuroglial cells using optimal induction medium,and the results were detected through the observation of morphology and special staining;ADMSCs were labeled by PKH26 in vitro,partially hepatectomized animal models were made and the labeled ADMSCs were autotransplanted by portal vein injection.Two weeks later,the livers were cut offand frozen sections were made,The labeled cells in the liver were observed by fluorescence microscope and their albumin expression was detected by immunofluorescence.Results: ADMSCs can be obtained from the adipose tissue and can be easily amplified in vitro,they can be induced into adipose cells,osteoblasts and neuroglial cells by using different inducing medium.Two weeks later,there were sporadic PKH26-labeled cells among hepatocytes in liver tissue section,and the immunofluorescence staining of albumin was positive.Conclusion:AMDSCs with multi-directional differ- entiation ability can be obtained from adipose tissue.ADMSCs can differentiate into hepatocytes in regenerative environment and may participate in liver regeneration.