碱性成纤维细胞生长因子-壳聚糖载体诱导神经干细胞分化及与肌细胞的共培养  

Basic fibroblast growth factor-chitosan carrier induces neural stem cells to differentiate into neurons followed by co-culture with myocytes

作　　者：翟婧妍 段红梅[1] 尚俊奎 杨朝阳[1,2] 李晓光[1,2] 

机构地区：[1]首都医科大学神经生物学系,北京市100069 [2]北京航空航天大学生物与医学工程学院,北京市100191

出　　处：《中国组织工程研究》2017年第6期877-882,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金国际合作与交流项目(31320103903);国家自然科学基金项目(31130022)~~

摘　　要：背景:神经干细胞具有自我更新和多向分化的潜能,高比例诱导其向神经元分化具有很大的应用前景。目的:探讨碱性成纤维细胞生长因子-壳聚糖载体体外对神经干细胞向神经元分化的影响,及分化的神经元与肌细胞共培养是否能形成突触样联系。方法:纯化后的神经干细胞分别与单纯壳聚糖、可溶性碱性成纤维细胞生长因子及碱性成纤维细胞生长因子-壳聚糖载体共培养。诱导7 d后,通过β-tubulinⅢ染色观察向神经元的分化;通过β-tubulinⅢ和Cha T染色观察向胆碱能神经元的分化;通过β-tubulinⅢ和MHC染色观察和肌细胞之间的突触联系。结果与结论:碱性成纤维细胞生长因子-壳聚糖载体组,分化形成神经元的比例是74%,显著高于其他2组,分化的神经元与共培养的肌细胞能形成突触样联系。提示碱性成纤维细胞生长因子-壳聚糖载体体外能够促进神经干细胞的分化,且分化形成的神经元能与共培养的肌细胞形成突触样联系。BACKGROUND： Neural stem cells（NSCs） hold self-renewal and multi-directional differentiation potential. NSCs differentiation into neurons in high proportion under induction conditions exhibits broad application prospect. OBJECTIVE： To explore the effect of basic fibroblast growth factor（b FGF）-chitosan carrier on the NSCs differentiation into neurons in vitro, and whether the differentiated neurons could form synaptic-like connection with myocytes. METHODS： After purification, the NSCs were co-cultured with chitosan, soluble b FGF or b FGF-chitosan carrier. After 7-day induction, the NSCs differentiation into neurons was observed by immunofluorescence staining of beta tubulin Ⅲ. The NSCs differentiation into cholinergic neurons was observed through double immunofluorescence staining of Cha T and beta tubulin Ⅲ. The synaptic-like connection between the neurons and myocytes was observed by triple staining of beta tubulin Ⅲ and MHC.RESULTS AND CONCLUSION： The percentage of differentiated neurons in the b FGF-chitosan carrier group was 74%, which was significantly higher than that in the other two groups. Additionally, the synaptic-like connection formed between the differentiated neurons and myocytes. To conclude, the b FGF-chitosan carrier promotes the NSCs differentiation into neurons to form synaptic-like connection with the co-cultured myocytes.

关 键 词：成纤维细胞生长因子2 神经元 组织工程 生物材料 材料相容性 碱性成纤维细胞生长因子 壳聚糖 神经干细胞 肌细胞 突触 共培养 国家自然科学基金 

分 类 号：R318[医药卫生—生物医学工程]