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目的:研究脐血间充质干细胞生物学特性及向神经元样细胞分化的潜能。方法:采用密度梯度离心结合贴壁培养法自脐血中分离间充质干细胞,观察细胞生长情况,描绘生长曲线,流式细胞仪检测细胞表面标志物,分别向成骨细胞、脂肪细胞、神经元样细胞进行诱导分化,通过茜素红染色、油红O染色检测脐血间充质干细胞成骨、成脂肪细胞诱导分化能力,而以免疫组织化学检测诱导后细胞表面神经标志物的表达。结果:纯化的脐血间充质干细胞贴壁生长,呈均一梭形,生长曲线呈S型,并以P3代增殖能力最强,细胞表面不表达或弱表达CD34、CD35、CD106,高表达CD29、CD44、CD105。成骨诱导2周后,可检测到钙化基质的形成,成脂肪诱导3周后,可检测到脂滴的形成。向神经元样细胞诱导分化后,可观察到典型的神经元样形态改变,且NSE、NF、GFAP阳性表达。结论:分离纯化的脐血间充质干细胞具有较强的增殖能力与分化潜能,并在体外诱导条件下可以向神经元样细胞定向分化。
Objective: To study the biological characteristics of cord blood mesenchymal stem cells and the potential of differentiating into neuron-like cells. METHODS: Mesenchymal stem cells (MSCs) were isolated from cord blood by density gradient centrifugation and adherent culture. The growth of cells was observed, and the growth curve was drawn. Cell surface markers were detected by flow cytometry. Osteoblasts, adipocytes and nerves The cells were induced to differentiate. Alizarin red staining and oil red O staining were used to detect the bone formation of umbilical cord blood mesenchymal stem cells. Adipocytes were induced to differentiate into adipocytes. The expression of neural cell surface markers was detected by immunohistochemistry. Results: The purified umbilical cord blood mesenchymal stem cells adherently grew in the shape of a uniform spindle with a S-shaped growth curve and the strongest proliferation ability in the P3 generation. The expression of CD34, CD35, CD106 and CD29 , CD44, CD105. After two weeks of osteogenic induction, formation of calcified matrix was detectable, and formation of lipid droplets was detectable 3 weeks after adipogenesis. After differentiation into neuron-like cells, typical neuronal-like morphological changes were observed, and NSE, NF and GFAP were positive. CONCLUSION: The purified cord blood MSCs have strong proliferation and differentiation potential and can be differentiated into neuron-like cells under in vitro induction conditions.