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| 大鼠外周血内皮祖细胞分离培养与鉴定 |
| Isolation, culture and identification of endothelial progenitor cells from peripheral blood of rats |
| 投稿时间：2017-04-16 |
| DOI：10.3969/j.issn.1000-0399.2017.10.001 |
| 中文关键词: 内皮祖细胞 细胞分离 细胞培养 细胞鉴定 |
| 英文关键词: Endothelial progenitor cells Cell isolation Cell culture Cellidentification |
| 基金项目:国家自然科学基金资助项目（项目编号：81171173）；安徽省自然科学基金资助项目（项目编号：11040606Q25） |
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| 中文摘要: |
| 目的 获得大鼠外周血内皮祖细胞（EPCs）并进行鉴定。方法 首先采用密度梯度离心法获得大鼠外周血单个核细胞，然后用EGM-2完全培养基体外培养条件下诱导单个核细胞分化为靶标细胞，最后分别用EPCs特异性标记物CD133和Flk-1检测及内吞乙酰低密度脂蛋白（ac-LDL）和荆豆凝集素-1（UEA-1）的能力，识别和鉴定靶标细胞。结果 靶标细胞培养至第6天呈纺锤形，梭状排列，具有与典型EPCs形态一致的生物学特征，CD133及FLK-1双抗体荧光检测结果阳性，内吞ac-LDL及UEA-1的能力实验结果阳性，证明该纺锤形细胞为EPCs。结论 通过密度梯度离心法及多细胞因子诱导大鼠外周血单个核细胞获得EPCs的方法可行，该方法可为EPCs的基础研究及临床应用提供理论与实验方法学基础。 |
| 英文摘要: |
| Objective To obtain and identify the endothelial progenitor cells (EPCs) from rat peripheral bloodusing density gradient centrifugation method. Methods The mononuclear cells were isolated from the peripheral blood of rats by density gradient centrifugation. The mononuclear cells were induced to differentiate into the target cells by EGM-2 complete medium. After that, the EPCs specific markers CD133 and Flk-1 were used to detect the target cells.Low density lipoprotein (ac-LDL) (Ac-LDL) and Ulex europaeus agglutinin-1 (UEA-1)were also used to detect the target cells.Results On day 6 of culture, target cells became spindle, spindle arrangement, consistent with the typical morphological characteristics of EPCs. CD133 and FLK-1 fluorescence detection showed positive result.In addition, endocytosis ac-LDL and UEA-1 test showed positive results, which proved that the spindle cells were EPCs.Conclusion It is feasible to obtain EPCs by density gradient centrifugation of peripheral blood mononuclear cells and the induction of multicellular factors. This method can provide the theoretical and experimental method for the basic research and clinical application of EPCs. |
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