脐血单个核细胞体外培养和诱导后神经干细胞标志物mRNA的表达  

Expression of neural stem cell-specific molecules cultured in vitro and induced mononuclear cells derived from human cord blood

作　　者：向静[1] 江德鹏[2] 王昌铭[1] 王景周[1] 

机构地区：[1]第三军医大学大坪医院野战外科研究所神经内科,重庆400042 [2]第三军医大学西南医院急诊科,重庆400038

出　　处：《重庆医科大学学报》2005年第3期352-355,401,共5页Journal of Chongqing Medical University

基　　金：重庆市科委攻关项目(7830)

摘　　要：目的:探讨人脐血单个核细胞(humancordbloodmonocytes,HCMNCs)培养增殖过程中的形态变化及诱导后神经干细胞标志物mRNA的表达。方法:采用OLYMPUS倒置显微镜观察培养过程中HCMNCs形态的变化;RT-PCR方法鉴定MNCs用神经生长因子(NGF)和维甲酸(RA)诱导前后神经干细胞(neuralstemcells,NSCs)标志物巢蛋白(Nestin)及Musashi-1蛋白mRNA的表达。结果:HCMNCs培养前,胞体较小,呈均一的圆形;培养后,细胞胞体增大,有破骨样细胞和间质样细胞两种形态。破骨样细胞胞体较大,呈圆形,多个核。间质样细胞胞体呈较均一的长梭形,形态类似于骨髓MSCs,但较骨髓MSCs稍小。RT-PCR检测提示,诱导前HCMNCs中Nestin弱表达,Musashi-1无表达;诱导后两者均表达增强,之后逐渐减弱。结论:使用密度梯度离心法可以从脐血中分离得到具有多向分化潜能的HCMNCs,经过EGF和RA诱导后,神经干细胞标志表达一过性增强后减弱。Objective:To investigate the configuration changes of human cord blood monocytes (HCMNCs)under specific culture conditions and the expression of NSCs marker under induction.Methods:HCMNCs were prepared from normal placenta after full term normal delivery. Observe the shape by microscope and reverse transcriptase polymerase chain reaction (RT-PCR) for nestin and musashi-1 antigen were performed to confirm nestin and musashi-1 antigen expression in HCMNCs.Results:RT-PCR demonstrated small amount of nestin expressed in HCMNCs but no musashi-1 antigen before induction, and both showed the strongest expression in 48h,then faded away gradually. The uncultivated MNCs were small and round.After culturing,the cells became larger,some cells change their shape into rhombic.Conclusion:The newly HCMNCs isolated by centrifugation over Lymphoprep densitgradient expressed nestin mRNA in low level and no musashi-1 show up,after induction,the former and musashi-1 increased intermittently and then decreased with the time of culturing.

关 键 词：脐血单个核细胞 巢蛋白 MUSASHI-1 RT—PCR 

分 类 号：R743[医药卫生—神经病学与精神病学]