悬浮生长的人脐血细胞中Nestin及MAP_2 mRNA的表达  

Nestin and MAP_2 mRNA expression in cultured nuadhensive human umbilical cord blood cells

作　　者：鄢文海[1] 刘计荣[2] 王建枝[1] 邢莹[3] 许燕[2] 韩雪飞[3] 曹孟德[3] 

机构地区：[1]华中科技大学同济医学院病理生理学教研室,武汉430030 [2]郑州大学基础医学院病理生理学教研室,郑州450052 [3]郑州大学医学院干细胞研究中心,郑州450052

出　　处：《郑州大学学报（医学版）》2004年第4期592-595,共4页Journal of Zhengzhou University(Medical Sciences)

基　　金：河南省重点科技攻关基金资助项目 0 1170 3 19;河南省医学科技创新人才工程项目 2 0 0 2 2 0 7

摘　　要：目的 :探讨悬浮生长的人脐血细胞中神经元特有标志物的表达特性 ,进一步阐明脐血单个核细胞(MNCs)向神经细胞分化的条件和机制。方法 :取健康自然分娩产妇脐血 ,密度梯度离心法分离单个核细胞 ,以 1×1 0 6ml-1 细胞密度接种于含体积分数为 2 0 %胎牛血清的DMEM培养瓶内 ,置 37℃、体积分数为 5 %CO2 、饱和湿度的培养箱内培养 ,倒置显微镜下观察细胞形态变化。收集培养第 1d、3d、5d、7d、1 0d悬浮生长的细胞 ,用RT PCR法检测培养前后MNCs神经元特有标志物Nestin和MAP2 的mRNA的表达。结果 :培养前MNCs呈小圆形 ,Nestin、MAP2mRNA均表达阳性。培养后细胞体积增大 ,部分细胞贴壁生长 ,部分细胞呈悬浮生长。收集悬浮生长细胞 ,第 1d、3dNestinmRNA的表达升高 ,随后降低直至转阴 ,而MAP2 mRNA的表达随培养时间的延长而增强。结论 :悬浮生长的脐血细胞表达神经元特有标志物Nestin和MAP2 mRNA 。Aim: To explore expression of Nestin and MAP 2 mRNA in cultured nuadhensive human umbilical cord cells and to further illustrate the environments and mechanism of mononuclear cells(MNC) from human umbilical cord blood different iating into neuron.Methods: MNC were iso lated from heparinized human umbilical cord(>50 ml)samples by centrifugation over Lymphoprep (density1.077 g/ml) densitgradient. MNC (1×10 6/ml cell density) were cultured in DMEM medium containing 20% (V/V)fetal bovine serum, at 37 ℃,5%CO 2,in a fully humidified atmosphere, and observed by using inverse phase contrast microscopy. On the day of 1,3,5,7,10, the nuadhensive cells were collected, Nestin and MAP 2 mRNA were determined by reverse transcription polymerase chain reaction(RT PCR). Results: The uncultivated MNCs were small and round. Nestin and MAP 2 mRNA were positively expressed. After culturing, the cells became larger, some cells were adhesive on the wall of the flask, but some cells were suspended growing. During culturing, the expression of Nestin mRNA were upregulated on the 1st and 3rd day, then declined and even not detectable on the 10th day; but the expression of MAP 2 mRNA was continuely upregulated during the period of culturing. Conclusion: Nuadhensive cells from human umbilical cord blood express Nestin and MAP 2 mRNA, the former decreases with the time of culturing but the later increased after culturing.

关 键 词：脐血细胞 神经干细胞 NESTIN 微管相关蛋白-2 

分 类 号：R331[医药卫生—人体生理学]