人脐血间充质干细胞体外向神经元样细胞分化的实验研究  

Experimental study on human umbilical cord blood mesenchymal stem cells induced into neuron-like cells

作　　者：杜江榕[1] 张军宏[1] 李继云[1] 江珊[1] 

机构地区：[1]北京大学深圳医院脊柱外科,广东省518036

出　　处：《实用诊断与治疗杂志》2007年第4期254-255,258,I0001,共4页Journal of Practical Diagnosis and Therapy

基　　金：深圳市科技计划项目(200603052)

摘　　要：目的:探讨来源于人脐带血的间充质干细胞体外诱导分化成神经元样细胞的可行性。方法:采用密度梯度离心方法分离人脐带血单个核细胞,用MesenCult培养液体外扩增生长传代培养,于倒置显微镜下观察其生长特性。取第3代的脐血间充质干细胞,用10μg／L成纤维细胞生长因子和30μmol／L全反式维甲酸诱导培养的细胞向神经细胞分化。倒置相差显微镜观察细胞形态变化,于诱导前、诱导后5 d和10 d分别进行神经元特异性烯醇化酶和胶质纤维酸性蛋白(GFAP)免疫细胞化学染色。结果:培养3代的脐血间充质干细胞经全反式维甲酸和成纤维细胞生长因子诱导后发生形态改变,呈双极或多极的神经元样形态,诱导5 d时神经元特异性烯醇化酶表达率为(30.3±5.2)％,而诱导10 d时达到(53．9±6.5)％,二者差异有统计学意义(P<0．05),不表达胶质纤维酸性蛋白。结论:脐血间充质干细胞经成纤维细胞生长因子和全反式维甲酸体外诱导,能够分化为神经元样细胞。Objective To investigate the feasibility of human umbilical cord blood mesenchymal stem cells induced into neuron-like cells. Methods Mononuclear cells were prepared from heparinized human umbilical cord blood samples by centrifugation over Lymphoprep and cultured in MesenCult medium. After treated with 30μmol/L retinoic acid and 10μg/L basic fibroblast growth factor, morphological changes of the cells were observed under phase contrast microscope. The cells were identified by immunocytochemistry with neuron specific enolase and glial fibrillary acidic protein antibodies respectively. Results Human umbilical cord blood mesenchymal stem cells were induced into neuron-like cells in a specific cell culture medium by retinoic acid and basic fibroblast growth factor. Neuron specific enolase was expressed （30.3±5.2）%after retinoic acid, and basic fibroblast growth factor treatment in 5 days but increased to （53.9±6.5）% in 10 days, There was a significant difference between 5 days and 10 days induction by retinoic acid and basic fibroblast growth factor （P〈0.05）. No glial fibrillary acidic protein was expressed in cells. Conclusion Human umbili blood mesenchymal stem cells are capable of differentiating into neuron-like-cells induced by acid and basic fibroblast growth factor in vitro.

关 键 词：脐带血 间充质干细胞 诱导 神经元样细胞 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]