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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。
OBJECTIVE: To obtain mesenchymal stem cells (ADMSCs) from adipose tissue and to verify their multidrug differentiation potential and to explore the role of ADMSCs in liver regeneration. Methods: The adipose tissue of rats was obtained. The stem cells were obtained by digestion with collagenase and amplified in vitro. The third passage cells were induced to osteoblast and adipogenesis with different inducing culture medium. After induced by cell morphology and Special staining observed induction effect. The cells were labeled with PKH26, and a partial hepatectomy model was made. The labeled autologous ADMSCs were implanted into the body through the portal vein. After 2 weeks, the liver was cut out to make frozen sections. The localization of the implanted cells in the liver was observed by fluorescence microscopy. Albumin expression. Results: The cells isolated from adipose tissue could be expanded in large quantities in vitro and could be induced to differentiate into osteoblasts and adipocytes. After 2 weeks ADMSCs were transplanted, PKH26 labeled cells scattered in the liver and immunofluorescent staining showed that Cell albumin staining positive. CONCLUSION: Mesenchymal stem cells with multi-directional differentiation potential can be obtained from adipose tissue of rats. The cells can differentiate into hepatocytes in liver regeneration environment and participate in liver regeneration.