| 兔骨髓源性血管内皮祖细胞的分离、培养及鉴定 |
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| 引用本文： | 吴波,卢正茂,王尧,罗天航,薛绪潮,毕建威,康俊升,方国恩.兔骨髓源性血管内皮祖细胞的分离、培养及鉴定[J].中国实验血液学杂志,2010,18(2):454-457. |
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| 作者姓名： | 吴波 卢正茂 王尧 罗天航 薛绪潮 毕建威 康俊升 方国恩 |
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| 作者单位： | 1. 第二军医大学附属长海医院普通外科,上海,200433
2. 解放军第88医院普通外科,山东泰安,271000 |
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| 基金项目： | 国家自然科学基金资助项目,编号30672170 |
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| 摘 要： | 本研究探讨兔骨髓源性血管内皮祖细胞(endothelial progenitor cells,EPC)的分离、培养及鉴定方法。抽取兔骨髓细胞,用梯度密度离心法获得单个核细胞,以内皮细胞培养液培养,通过细胞形态观察、免疫组织化学试验、流式细胞术以及内皮祖细胞吞噬功能进行鉴定。结果表明,新分离的骨髓单个核细胞呈圆形,培养48小时后可见贴壁细胞呈集落样生长,细胞呈圆形或不规则形,核分裂相明显,至培养第7天成片生长的细胞集落相互连接呈梭形的内皮样细胞。内皮祖细胞免疫组织化学检测结果显示CD133(+),CD34(+),Ⅷ因子(++),KDR(++);流式细胞术鉴定结果显示CD133的阳性率为(18.23±7.12)%,CD34的阳性率为(47.71±14.85)%,CD31的阳性率为(71.61±13.51)%,KDR的阳性率为(87.24±11.40)%。细胞吞噬功能鉴定说明超过80%的贴壁细胞都特异性地摄取了Dil-acLDL和FITC-UEA-1。结论:密度梯度离心法体外分离兔骨髓源的单个核细胞,在一定的诱导培养条件下能分化成为血管内皮祖细胞。
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| 关 键 词： | 骨髓 内皮祖细胞 细胞培养 细胞分离 细胞鉴定 |
Isolation, Culture and Identification of Rabbit Bone Marrow-derived Vascular Endothelial Progenitor Cells |
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| WU Bo,LU Zheng-Mao,WANG Yao,LUO Tian-Hang,XUE Xu-Chao,BI Jian-Wei,KANG Jun-Sheng,FANG Guo-En.Isolation, Culture and Identification of Rabbit Bone Marrow-derived Vascular Endothelial Progenitor Cells[J].Journal of Experimental Hematology,2010,18(2):454-457. |
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| Authors: | WU Bo LU Zheng-Mao WANG Yao LUO Tian-Hang XUE Xu-Chao BI Jian-Wei KANG Jun-Sheng FANG Guo-En |
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| Institution: | WU Bo,LU Zheng-Mao,WANG Yao,LUO Tian-Hang,XUE Xu-Chao,BI Jian-Wei,KANG Jun-Sheng1,FANG Guo-En Department of General Surgery,Changhai Hospital,The Second Military Medical University,Shanghai,China,1Department of General Surgery,Chinese PLA Hospital No88,Taian,271000,Sh,ong Province |
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| Abstract: | The aim of study was to set up a suitable method of isolation,culture and identification of endothelial progenitor cells(EPC)derived from rabbit bone marrow.Density gradient centrifugation was used to isolate mononuclear cells from bone marrow,the isolated mononuclear cells were cultured with specific culture medium for EPCs.EPCs were identified by cellular morphologic observation,immunohistochemistry testing,flow cytometry and the function test of taking up Dil-ac-LDL and FITC-UEA-1.The results indicated t... |
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| Keywords: | bone marrow endothelial progenitor cell cell culture cell isolation cell identification |
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