胎盘间充质干细胞体外分离培养及向成骨细胞和神经细胞分化的研究

发布时间：2018-07-20 22:05

【摘要】：目的：人足月胎盘间充质干细胞(hPMSCs)在体外进行分离纯化并对其生物学特征进行研究,建立共培养体系(分别与成骨细胞和神经细胞),观察共培养体系对hPMSCs分化为成骨细胞和神经细胞的影响。方法：采用胶原酶消化法从人胎盘中分离纯化间充质干细胞(MSCs);通过测细胞生长曲线、CD分子、细胞超微结构及成骨能力对hPMSCs进行鉴定；Transwell双层培养皿的底层接种成骨细胞或神经细胞,上层种hPMSCs接,加入高糖DMEM (10%胎牛血清),对照组上下层均接种hPMSCs。诱导后的成骨细胞用碱性磷酸酶及茜素红染色进行鉴定,诱导后的神经细胞用NSE进行鉴定。结果：培养14d后大量细胞贴壁并呈成纤维状生长；强表达CD44,CD29,不表达CD34,CD45;诱导后可以向成骨细胞分化；与成骨细胞的共培养组碱性磷酸酶活性染色阳性,对照组碱性磷酸酶活性染色成阴性。与神经细胞共培养的细胞可见丝状突起,成细长型,NSE染色为阳性。结论：MSCs同样可见于人足月胎盘中,其生物学特性与其他来源的MSCs相似,成骨细胞及神经细胞生长过程中提供的微环境可以诱导hPMSCs向成骨细胞及神经细胞分化。
[Abstract]:Objective: to isolate and purify human full-term placental mesenchymal stem cells (hPMSCs) in vitro and study their biological characteristics. The effects of co-culture system on differentiation of hPMSCs into osteoblasts and neural cells were observed. Methods: mesenchymal stem cells (MSCs) were isolated and purified from human placenta by collagenase digestion. The bottom layer of Transwell culture dish was inoculated with osteoblasts or nerve cells, the upper layer of HPMSCs was added with high glucose DMEM (10% fetal bovine serum), and the upper and lower layers of control group were inoculated with hPMSCs. The induced osteoblasts were identified by alkaline phosphatase and alizarin red staining, and the induced neurons were identified by NSE. Results: after 14 days of culture, a large number of cells adhered to the wall and showed fibrous growth, strong expression of CD44 + CD29, no expression of CD34 + CD45, differentiation into osteoblasts after induction, and positive staining of alkaline phosphatase activity in co-culture group with osteoblasts. Alkaline phosphatase activity in control group was negative. The cells co-cultured with nerve cells showed filamentous processes and were positive for NSE staining. Conclusion\%\
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R329

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