贵州医科大学学报

2020, v.45;No.234(03) 265-269

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大鼠骨髓源内皮祖细胞的分离与鉴定
Isolation and Identification of Endothelial Progenitor Cells from Rat Bone Marrow

Nganguem Nzalle Yranney Brice;林慕之;毛善永;张蓓;周海燕;胡柏龙;王艺明;况春燕;刘兴德;
Nganguem Nzalle Yranney Brice;LIN Muzhi;MAO Shanyong;ZHANG Bei;ZHOU Haiyan;HU Bailong;WANG Yiming;KUANG Chunyan;LIU Xingde;Department of Cardiovascular Medicine,the Affiliated Hospital of Guizhou Medical University;Department of Cardiovascular Medicine,Guizhou Provincial People's Hospital;Department of Ultrasound,the Affiliated Hospital of Guizhou Medical University;Department of Anesthesiology,the Affiliated Hospital of Guizhou Medical University;Department of Psychology,the Affiliated Hospital

摘要(Abstract):

目的:建立大鼠骨髓源内皮祖细胞(EPCs)的分离与鉴定方法。方法:取大鼠双侧股骨和胫骨骨髓腔细胞,采用密度梯度离心法、组织淋巴细胞分离液分离单个核细胞,将细胞过滤离心后置于含20%FBS的DMEM低糖完全培养基中培养;采用流式细胞仪鉴定EPCs细胞表面标志物CD34、CD133及VEGFR2的表达。结果:细胞培养2 d后可观察到梭形的贴壁细胞,培养7 d后细胞呈克隆样集落,流式细胞仪鉴定为EPCs细胞的特征。结论:大鼠骨髓中可提取并分化出质量较好的EPCs细胞。
Objective: To establish a method to isolate and identify rat bone marrow-derived endothelial progenitor cells( EPCs). Methods: Rat femoral and tibia bone marrow cells were collected,and mononuclear cells were separated by density gradient centrifugation and tissue lymphocyte separation solution. The cells were filtered,centrifuged and cultured in DMEM with low glucose supplemented with 20% fetal bovine serum. The expression levels of CD34,CD133 and VEGFR2 on the surface of EPCs were measured by flow cytometry. Results: The spindle-shaped adherent cells were observed after 2 days of cell culture,and the cells showed colony-like colonies after7 days of culture. The characteristics of EPCs cells were identified by flow cytometry. Conclusion:EPCs with good quality can be extracted and differentiated from rat bone marrow.

关键词(KeyWords): 大鼠;骨髓细胞;内皮祖细胞;分离;鉴定
rats;bone marrow cells;endothelial progenitor cells(EPC);isolation;identification

Abstract:

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基金项目(Foundation): 国家自然科学基金(31760294,81560056,81904319,81960315);; 贵州省科技厅基金[黔科合基础(2016)1120],[黔科合平台人才(2018)5608],[黔科合平台人才(2018)5779-36],[黔科合平台人才(2018)5779-52];; 贵阳市科技局基金[筑科合同(2017)5-14],[筑科合同(2017)30-10];; 贵州省卫生计生委基金[gzwjkj(2017-1-016)];; 贵州省教育厅青年科技人才成长项目[黔教合KY字(2018)182];; 贵州省人社厅留学人员科技创新项目[黔人项目资助合同(2018)0003]

作者(Author): Nganguem Nzalle Yranney Brice;林慕之;毛善永;张蓓;周海燕;胡柏龙;王艺明;况春燕;刘兴德;
Nganguem Nzalle Yranney Brice;LIN Muzhi;MAO Shanyong;ZHANG Bei;ZHOU Haiyan;HU Bailong;WANG Yiming;KUANG Chunyan;LIU Xingde;Department of Cardiovascular Medicine,the Affiliated Hospital of Guizhou Medical University;Department of Cardiovascular Medicine,Guizhou Provincial People's Hospital;Department of Ultrasound,the Affiliated Hospital of Guizhou Medical University;Department of Anesthesiology,the Affiliated Hospital of Guizhou Medical University;Department of Psychology,the Affiliated Hospital

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DOI: 10.19367/j.cnki.1000-2707.2020.03.004

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