体外扩增巨核细胞的研究

发布时间：2018-01-05 17:29

本文关键词：体外扩增巨核细胞的研究　出处：《中国协和医科大学》2009年硕士论文　论文类型：学位论文

【摘要】： 目的:探讨采用脐血单个核细胞为起始细胞在无血清培养基中体外扩增巨核细胞;观察胎儿肺部组织来源的间充质干细胞对脐血单个核细胞诱导分化为巨核细胞的影响。方法:分取脐血单个核细胞,在TPO,IL—11,肝素的扩增体系中分为两组培养,第一组以CD34~+细胞作为对照,单独培养,第二组与间充质干细胞共培养,在7天,10天,14天时,取细胞进行活细胞计数,流式细胞仪分析CD41a和CD61的表达情况。取14天细胞采用免疫组化和透射电镜分析细胞形态与结构,通过流式细胞仪分析细胞DNA含量。结果:MNC经细胞因子诱导后,CD41a和CD61阳性比例明显上升,与0天时相比,培养14天后,CD41a~+细胞平均扩增了429.5倍,CD61~+细胞平均扩增了339倍。第二组在10天时得到的CD41~+细胞和CD61~+细胞最多,分别是第一组的4.5倍和4.7倍,免疫组化和电镜结果表明,两组细胞核发育不成熟。结论:在TPO,IL—11,肝素的扩增体系下以MNC为起始细胞能够诱导得到大量的巨核细胞,胎肺间充质干细胞能有效刺激CD41~+细胞和CD61~+细胞生成,对细胞核发育影响。
[Abstract]:Objective: to investigate the expansion of megakaryocytes from cord blood mononuclear cells in serum-free medium in vitro. To observe the effect of mesenchymal stem cells derived from fetal lung tissue on the differentiation of cord blood mononuclear cells into megakaryocyte. The amplification system of heparin was divided into two groups. The first group was cultured alone with CD34 ~ cells and the second group co-cultured with mesenchymal stem cells. The living cells were counted and the expression of CD41a and CD61 were analyzed by flow cytometry. The morphology and structure of the cells were analyzed by immunohistochemistry and transmission electron microscopy on the 14th day. Results the positive rates of CD41a and CD61 were significantly increased after induced by cytokines. Compared with 0 days, the positive rates of CD41a and CD61 were significantly increased after 14 days of culture. CD41a~ cells were amplified by an average of 429.5 times and CD61- cells by 339 times on average. In the second group, the number of CD41 ~ cells and CD61 ~ cells were the highest at 10 days. The results of immunohistochemistry and electron microscope showed that the nuclei of the two groups were immature. In the system of heparin amplification, a large number of megakaryocytes could be induced by using MNC as the starting cells, while fetal lung mesenchymal stem cells could effectively stimulate the generation of CD41 ~ cells and CD61 ~ cells. Effect on nuclear development.
【学位授予单位】：中国协和医科大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R329

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