摘要
目的:探讨人脐带间充质干细胞(h UC-MSCs)体外定向分化为神经元细胞的适宜条件,提高h UC-MSCs的分化效率,为神经细胞移植提供新细胞来源。方法:采用组织贴壁法分离培养h UC-MSCs,并应用流式细胞术鉴定其表面抗原;利用经典诱导(方案1)和复合诱导(方案2)两种方法定向诱导h UC-MSCs分化为神经元细胞,并以只加培养基的h UC-MSCs作对照。倒置显微镜下观察细胞形态变化,免疫细胞化学检测神经元迁移蛋白(DCX)、神经元特异性烯醇化酶(NSE)的表达,qRT-PCR检测各组神经分化标记基因DCX、NSE、Mash、Ngn1的转录水平。结果:与对照组相比,两种方案均能诱导hU C-MSCs分化为神经元样细胞,开始表达DCX和NSE;但是,与方案1相比,方案2组的细胞NSE的阳性表达率更高(P<0.05),各神经分化标记基因mRNA表达水平升高(P<0.05),向神经元样细胞分化更明显。结论:复合诱导方案能有效改善神经分化微环境,促进hU C-MSCs定向分化为神经元样细胞。
Aim： To optimize the culture conditions for human umbilical cord mesenchymal stem cell（ h UC-MSCs）and increase the differentiation efficiency. Methods： The h UC-MSCs were isolated form human umbilical cord by tissue adherence and its surface antigens were identified by flow cytometry. Classic strategy and composite strategy were used to induce the differentiation of h UC-MSCs into neurons,and the cells were randomly allocated into three groups： control group,classic strategy group（ group 1） and composite strategy group（ group 2）. The morphology,the DCX / NSE positive cell ratio and expressions of neuronal differentiation genes DCX,NSE,Mash and Ngn1 were analyzed by microscope,immunohistochemistry and qRT-PCR after induced for 4 d in different groups,respectively. Results： Compared with control group,the group 1 and group 2 in vitro could both induce the neural differentiation of h UC-MSCs,and the expressions of neuronal differentiation proteins DCX and NSE could be detected. Compared with group 1,composite microenvironment demonstrated a higher NSE positive ratio（ P 0. 05） and mRNA expressions of neural differentiation mark genes were also higher（ P 0. 05）,leading to differentiation towards neuron-like cells more likely. Conclusion： The composite strategy could significantly improve neural differentiation microenvironment,promote h UC-MSCs differentiation towards neuron-like cells.
出处
《郑州大学学报（医学版）》
CAS
北大核心
2016年第4期469-474,共6页
Journal of Zhengzhou University(Medical Sciences)
基金
国家自然科学基金资助项目81471306
U1404313
河南省科技创新人才计划154200510008
河南省高校科技创新团队支持计划15IRTSTHN022
河南省产学研合作项目142107000008
