摘要
背景:目前关于骨髓间质干细胞能否向神经元方向分化的报道不多,且争论多集中在分化后的神经元是否仅具有神经元形态而不具有神经元功能。目的:探讨海马神经元条件培养液诱导大鼠骨髓间质干细胞向神经元样细胞和神经胶质样细胞分化的可能性。方法:将第5代大鼠骨髓间质干细胞分为4组:条件培养基组加入海马神经元和胶质细胞的培养液;b-FGF组加入含b-FGF的DMEM培养基;无血清培养组加入含Neurobasal和B27的无血清培养基;阴性对照组加入含胎牛血清的DMEM。各组诱导12,24h后,应用免疫细胞化学染色行神经元特异性烯醇化酶、微管相关蛋白2、胶质纤维酸性蛋白的鉴定,Western-blot法检测细胞神经元特异性烯醇化酶、微管相关蛋白2和胶质纤维酸性蛋白的表达。结果与结论:诱导12,24h后,条件培养基组、b-FGF组、无血清培养组骨髓间质充干细胞微管相关蛋白2、胶质纤维酸性蛋白、神经元特异性烯醇化酶均呈阳性表达,阴性对照组未见表达。与阴性对照组比较,诱导后24h,条件培养基组、b-FGF组、无血清培养组微管相关蛋白2表达均明显增强(P<0.05),且条件培养基组增强幅度显著高于另两组(P<0.05);条件培养基组、b-FGF组、无血清培养组神经元特异性烯醇化酶及胶质纤维酸性蛋白表达无明显差异。结果证实海马神经元条件培养液可体外诱导大鼠骨髓间质干细胞分化为神经元样细胞和神经胶质样细胞,与含b-FGF的培养基和无血清培养基相比,海马神经元条件培养基诱导的神经元和神经胶质细胞阳性率最高。
BACKGROUND：There are few reports addressing the differentiation of bone marrow mesenchymal stem cells （BMSCs） into neurons,and the uncertainties mainly focused on the differentiated neurons had neuron morphology,but did not have neuron function.OBJECTIVE：To investigate the feasibility of rat bone marrow mesenchyma stem cells （BMSCs） differentiation into neuron-like cells and glial-like cells under rat hippocampal neuron＇s conditional medium.METHODS：Rat BMSCs at passage 5 were divided into 4 groups.The medium of hippocampal neurons and glial cells was added in the conditioned medium group.The Dulbecco＇s modified Eagle＇s medium containing bFGF was added in the basic fibroblast growth factor （bFGF） group.The serum-free medium containing Neurobasal and B27 was added in the serum-free medium group.The DMEM supplemented with fetal bovine serum was added in the negative control group.12 and 24 hours following induction,neuron specific enolase （NSE）,microtubule-associated protein-2 （MAP-2） and glial fibrillary acidic protein （GFAP） were detected using immunocytochemical staining in each group.NSE,MAP-2 and GFAP expression was determined using Western-blot assay.RESULTS AND CONCLUSION：12 and 24 hours following induction,BMSCs were positive for MAP-2,GFAP and NSE in the conditioned medium,bFGF and serum-free medium groups,but negative in the negative control group.Compared with the negative control group,MAP-2 expression was significantly enhanced in the conditioned medium,bFGF and serum-free medium groups 24 hours following induction （P 0.05）,and the increased range was significantly greater in the conditioned medium group compared with other two groups （P 0.05）.No significant difference in NSE and GFAP expression was detected in the conditioned medium,bFGF and serum-free medium groups.Results suggested that hippocampal neuron conditioned medium can in vitro induce the differentiation of rat BMSCs into neuron-like cells and glial cell-like cells.Compared with the bFGF medium and serum-free medium,positive rate was greatest in the hippocampal neuron conditioned medium-induced neurons and glial cells.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2010年第6期1105-1110,共6页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
the Natural Science Foundation of Hebei Province,No.C2009001081~~
