人诱导多能干细胞向外周神经元分化的实验研究  

Derivation of Peripheral Neurons from Human Induced Pluripotent Stem Cells

作　　者：林婉颐 黄利华[1] 周立文[1] 石翾 赖兴强[1] 李伟强[1] 

机构地区：[1]中山大学干细胞与组织工程研究中心,广东广州510080

出　　处：《中山大学学报（医学科学版）》2014年第3期431-437,共7页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金(81271265)

摘　　要：【目的】探讨人诱导多能干细胞(hiPS细胞)在体外向外周神经元分化的能力。【方法】人诱导多能干细胞在无饲养层培养条件下维持培养,然后在神经培养基中悬浮培养分化为神经球,通过把神经球贴壁在多聚赖氨酸和层粘连蛋白包被的培养板上,利用流式分选细胞方法获得神经嵴干细胞(NCSC),通过把神经嵴干细胞在外周神经诱导培养基中诱导分化,获得外周神经元,利用免疫荧光染色的方法检测神经嵴干细胞和外周神经元的标记物。【结果】贴壁的神经球出现特征性的神经花环(Neural rosettes)结构,神经嵴干细胞从神经花环发生迁移;神经嵴干细胞向外周神经元诱导后出现神经丝结构,并表达外周神经元特异性标记物。【结论】人诱导多能干细胞在体外能高效向外周神经元分化。[Objective] To discuss peripheral neurons differentiation ability of human induced pluripotent stem cells.[Methods] The human induced pluripotent stem cells were maintained under feeder-free condition.Then hiPS cells can give rise to neurospheres in suspension culture with neural culture medium.When neurospheres were replated on the poly-L-lysine/laminin-coated dish,the neural crest stem cells emerged and were enriched by FACS.Then the neural crest stem cells differentiate into peripheral neurons,which can be identified by immunocytochemical assays.[Results] When neurospheres were cultured under adherent condition,the neural rosettes emerged and a great amount of neural crest stem cells migrated from the rosette structures.The peripheral neurons were induced from the neural crest stem cells and expressed the special markers.[Conclusion] hiPS cells were successfully induced to peripheral neurons.

关 键 词：人诱导多能干细胞 外周神经元 神经嵴干细胞 

分 类 号：Q254[生物学—细胞生物学]