人脐带间充质干细胞体外扩增和向神经元样细胞定向诱导分化的研究

目的探讨人脐带间充质干细胞(MSCs)的体外分离、纯化、扩增和向神经元样细胞的定向诱导分化,以期为脐带MSCs的神经移植提供理论依据。方法无菌条件下收集剖宫产新生儿脐带,酶消化法获取MSCs,进行培养。用流式细胞仪检测MSCs的表面标志。取扩增3,5,10代的MSCs分别向神经元样细胞诱导,用免疫组化和RT-PCR法检测神经元样细胞特异性标志。结果脐带富含MSCs,且脐带MSCs(UCMSCs)强表达CD13、29、CD44、CD105,弱表达CD106,不表达CD34、CD11a、CD14、CD33、CD45。神经条件培养基诱导后的细胞平均有70%左右呈现典型的神经元样表型。免疫组化法检测发现不同代数的MSCs经诱导后均表达nestin,NSE,NeuN,NF-M,弱表达GFAP。RT-PCR显示诱导后NSEmRNA表达增加。结论MSCs存在于人脐带中,并且在体外有较强的增殖能力,特定条件下能够分化为神经元样细胞。

Amplification and differentiation towards neuron-like cells of human umbilical cord derived mesenchymal stem cells

Objective To explore the isolation, purification, amplification and differentiation towards neuron-like cells of human umbilical cord (UC) derived mesenchymal stem cells (MSCs) in order to offer some evidence for the transplantation of MSCs derived from human UC. Methods The UC from full-term neonates delivered abdominally was collected in sterile condition. MSCs were obtained by enzyme digestion and cultured in DMEM/F12 medium. Surface antigen of UC-derived MSCs was detected by flow cytometry. Different passages of MSCs (p2, p5, p10) were induced to differentiate towards neuron-like cells and then neural markers were detected by immunocytochemistry and RT-PCR analysis. Results UC is a rich source of MSCs, and MSCs expressed strongly CD13, CD29, CD44, CD105, weakly CD106, did not express CD34, CD11a, CD14, CD33, CD45. Under the induction of DMEM/F12 medium, about 70% cells rapidly assumed the morphology of multipolar neurons. Immunocytochemical assay showed that different passages of MSCs expressed nestin, NSE, NeuN, NF-M and weakly expressed GFAP after differentiation. RT-PCR also showed an increased NSE mRNA expression after induction. Conclusion UC-derived MSCs have a strong proliferation capacity and can be induced to differentiate towards neuron-like cells in vitro.