脐血单个核细胞培养前后神经细胞特有标志物的变化

摘要

背景:目前神经于细胞(Neural Stem Cells,NSCs)的来源仅限于胚胎,因此寻找一种新的NSCs的来源对于神经科学领域基础与应用研究的发展至关重要,而脐血中资源丰富.目的:探讨脐血单个核细胞培养前后神经细胞特有标志物的表达差异.地点与材料:研究地点为郑州大学医学院干细胞研究中心.材料为用肝素抗凝采血袋从郑州大学第一附属医院及第三附属医院产科无菌收集健康产妇足月孕娠顺产或剖宫产的要儿脐带血50～100 mL,充分混匀后,置保温盒中带回实验室.设计与方法:采用RT-PCR方法对脐血单个核细胞培养前后nestin,NF-M及MAP2 mRNA的表达进行检测.主要观察指标:脐血单个核细胞培养前后nestin,NF-M和MAP2 mRNA的表达.结果:nestin,NF-M及MAP2 mRNA在培养前后的脐血单个核细胞中表达呈阳性;同未培养的脐血单个核细胞相比,nestin,NF-M及MAP2mRNA在培养的脐血单个核细胞表达增强.结论:在直接分离的脐血单个核细胞中存在着神经(干)细胞;培养后,神经元样细胞增多,nesth,NF-M及MAP2 mRNA表达增强.%BACKGROUND: The source of neural stem cells(NSCs) now just limits tothe embryos so that it is extremely important to seek a new resource of NSCSfor further developing both foundational and applied research in the field ofneuroscience. Cord blood is found a good source of NSCs.OBJECTIVE: To explore the expression difference on specific markers ofcord blood mononuclear cells(MNCs) before and after culture.SETTING and MATERIALS: The research was done in the Stem CellResearch Center in the Medical College of Zhengzhou University. 50 - 100mL cord blood sample was obtained aseptically from infants delivered by fullterm normal labor or cesarean in Obstetrical Department of First and ThirdAffiliated Hospitals by using blood collection bags containing anticoagulantheparin. After fully mixed, the blood sample was transported to laboratory in athermal insulation box.DESIGN and METHODS: Expressions of Nestin, NF-M, MAP2 and mRNAwere assayed by RT-PCR method before and after culture of cord bloodMNCs.MAIN OUTCOME MEASURES: Expressions of Nestin, NF-M, MAP2 andmRNA before and after the culture of cord blood MNCs.RESULTS: There were positive expressions of Nestin, NF-M, MAP2 andmRNA both before and after the culture of cord blood mononuclear cells.Compared with non-cultured MNCs, the expressions of Nestin, NF-M, MAP2and mRNA were enhanced in the cultured sample.CONCLUSION: There were neural stem cells existing in the directly iso-lated cord blood mononuclear cells. Afrer culture, the expressions of Nestin,NF-M, MAP2 and mRNA were enhanced because of the increasing number ofneuron-like cells.