脐血单个核细胞体外向神经干细胞的诱导及Foxg1基因的表达  

Induction and Foxg1 gene expression of the neural stem cells from human umbilical cord blood mononuclear cells in vitro

作　　者：范亚珍[1] 王军[1] 陈海[1] 朱登纳[1] 吴值荣[1] 侯艳艳[1] 

机构地区：[1]郑州大学第三附属医院,郑州450052

出　　处：《山东医药》2010年第11期13-15,共3页Shandong Medical Journal

基　　金：河南省医学科技攻关计划项目(200703066)

摘　　要：目的探讨人脐血单个核细胞(UBC-MNCs)体外向神经干细胞(NSCs)的诱导分化机制及NSCs中Foxg1基因表达的意义。方法从脐血中分离出UBC-MNCs,用含hEGF、bFGF和B27因子的Neurobasal培养基联合诱导其向NSCs方向分化,观察NSCs形态学及增殖分化特点;免疫组化法检测培养细胞中神经细胞标志抗原Nestin、NSE、GFAP的表达情况,BrdU标记靶细胞;RT-PCR法检测分离后、培养3、6、9、12 d细胞中Foxg1基因的表达。结果UBC-MNCs可定向诱导分化为神经元样细胞,表达Nestin、NSE、GFAP标记抗原。Foxg1 mRNA在诱导前细胞中低表达,诱导后逐渐升高(P<0.05),6 d达高峰,之后逐渐下降(P<0.05)。结论体外定向诱导培养可获得UBC-MNCs源性NSCs,Foxg1基因是NSCs增殖分化关键调控因子,脐血可成为NSCs的新来源。Objective To explore the mechanism of human umbilical cord blood mononuclear cells （UBC-MNCs） inducing and differentiating into neural stem cells （NSCs） in vitro and the significance of Foxgl gene expression. Methods The UBC-MNCs were isolated from human umbilical cord blood, induced and diferentiated into NSCs in Neurobasai medium containing hEGF, bFGF and B27 faetures in vitro, the characteristics of morphology and proliferation, differentiation were observed ; the expression of specific neuron antigen markers of Nestin, NSE, GFAP were detected by immunohistochemlstry, target cells were traced by BrdU; the Foxgl gene expression was detected by RT-PCR at separation times of 3,6,9,12ds after culture. Results UBC-MNCs differentiated into neuron-like cells after orientid induction, and labelled antigen of Nestin, NSE and GFAP were expressed . Foxgl gene mRNA expressed lowlyat first, and increased gradually after the induction （ P 〈 0.05 ）, came to its peak at 6 days, then gradually declined （ P 〈 0.05 ）. Conclusions NSCs from UBC-MNCs can be obtained by targeting induced culture in vitro, Foxgl gene is the key regulation factor for the proliferation and diferentiation of NSCs, human cord blood can be the new source of NSCs.

关 键 词：Foxg1基因 脐血 神经干细胞 细胞培养 诱导分化 

分 类 号：R322.2[医药卫生—人体解剖和组织胚胎学] R714.5[医药卫生—基础医学]