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人脐血间充质干细胞(MSCs)是从脐带血中分离和培养的一种多潜能成体干细胞。本实验通过密度梯度离心法分离人脐血单个核细胞,贴壁法筛选出脐血MSCs,体外扩增,免疫荧光方法检测表面抗原。传3代的脐血MSCs经20ng/mlbFGF预诱导24h、3μg/μl丹参素诱导6h、20ng/mlEGF和20ng/mlbFGF诱导分化5d后出现类似神经元样细胞的形态改变,伸出长突起。免疫组织化学和免疫荧光方法鉴定显示,诱导后的细胞能特异性表达神经元特异性标志NeuN和β-TubulinⅢ,而星形胶质细胞特异性标志GFAP阳性细胞较少。上述结果表明,丹参素联合生长因子EGF和bFGF在体外可定向诱导人脐血MSCs分化为神经元样细胞。
Human umbilical cord blood mesenchymal stem cells (MSCs) are pluripotent adult stem cells isolated and cultured from umbilical cord blood. In this study, human umbilical cord blood mononuclear cells were isolated by density gradient centrifugation. Cord blood MSCs were screened by adherent method and amplified in vitro. The surface antigens were detected by immunofluorescence. The first generation of cord blood MSCs pre-induced by 20ng / mlbFGF 24h, 3μg / μl Danshensu induced 6h, 20ng / mlEGF and 20ng / mlbFGF induced differentiation 5d after the emergence of neuronal-like cells morphological changes, protruding long process. Immunohistochemistry and immunofluorescence assay showed that the induced cells specifically expressed neuron-specific markers NeuN and β-TubulinⅢ, while astrocyte-specific markers of GFAP-positive cells were less. The above results show that Danshensu combined with EGF and bFGF can induce MSCs differentiated into neuron-like cells in vitro.