大鼠自体外周血内皮前体细胞培养的实验研究

发布时间：2018-03-05 07:12

本文选题：细胞培养　切入点：外周血　出处：《青岛大学》2008年硕士论文　论文类型：学位论文

【摘要】： 目的探讨自大鼠外周血获取内皮前体细胞(endothelial progenitorcells EPCS)的方法,为干细胞移植治疗缺血性心肌损伤提供新的无创性的细胞来源。方法抽取经连续皮下注射大鼠粒细胞集落刺激因子(Rat GranulocyteColony Stimulating Factor rG-CSF)七天的成年雄性Sprague-Dawley(SD)大鼠外周血,应用聚蔗糖(Ficoll 400)密度梯度离心法分离提取大鼠外周血中的单个核细胞,然后以20%低糖IMDM、胎牛血清、马血清、碱性成纤维细胞生长因子(basic fibroblast growth factor bFGF)和血管内皮生长因子(vascular endothelial growth factor VEGF)等混合而成的完全培养液,进行体外培养扩增,培养所得细胞经CD31、CD34、Flk-1和vW F免疫荧光染色并检测其结合UEA-1、摄取ac-LDL的能力。结果培养所得细胞经CD31、CD34、Flk-1和vW F免疫荧光染色,并测定其结合UEA-1、摄取ac-LDL的能力,皆为阳性,证实为内皮前体细胞(EPC)。结论从粒细胞动员后的大鼠外周血中获取单个核细胞,经体外培养诱导,成功获得内皮前体细胞(EPC),内皮前体细胞(EPC)有望成为移植治疗缺血性心肌损伤的新的无创性细胞来源。
[Abstract]:Objective to investigate the method of acquiring endothelial progenitor cells (progenitorcells) from peripheral blood of rats, and to provide a new noninvasive cell source for stem cell transplantation in the treatment of ischemic myocardial injury. Methods the peripheral blood of adult male Sprague-Dawley SD rats injected subcutaneously with rat GranulocyteColony Stimulating Factor rG-CSF for 7 days was isolated and extracted by density gradient centrifugation with polysucrose Ficoll 400 (Ficoll 400), and the mononuclear cells were isolated from the peripheral blood of adult male Sprague-Dawley rats by density gradient centrifugation. Then, 20% low glucose IMDM, fetal bovine serum, horse serum, basic fibroblast growth factor basic fibroblast growth factor bFGFand vascular endothelial growth factor VEGF were mixed and cultured in vitro. The cultured cells were stained with CD31, CD34, Flk-1 and VWF, and the ability of UEA-1 to absorb ac-LDL was detected. Results the cultured cells were stained with CD31T CD34Flk-1 and VWF, and the ability of uptake of ac-LDL was positive, which was confirmed to be endothelial progenitor cells. Conclusion after granulocyte mobilization, mononuclear cells were obtained from peripheral blood of rats. After cultured and induced in vitro, endothelial progenitor cells (EPCs) were successfully obtained. EPCs of endothelial progenitor cells (EPCs) are expected to become a new noninvasive cell source for the treatment of ischemic myocardial injury.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R329;R541.4

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