促红细胞生成素对大鼠内皮祖细胞生物学特点的影响  

Effects of erythropoietin on biological characteristics of rat endothelial progenitor cells

作　　者：王东[1] 王亮[1] 刘庆国[2] 王晓楠[1] 尉辉杰[1] 张建宁[1] 

机构地区：[1]天津医科大学总医院神经外科,天津市300052 [2]山西长治医学院附属和平医院神经外科,山西省长治市047100

出　　处：《中国组织工程研究与临床康复》2010年第15期2705-2708,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金资助项目(30772229);国家教育部高等学校博士学科点专项科研基金(20070062008)~~

摘　　要：背景:内皮祖细胞是内皮细胞的前体细胞,可参与新生血管的形成。研究发现,促红细胞生成素对骨髓内皮祖细胞有动员作用,促进血管的新生和损伤组织修复。目的:观察不同浓度促红细胞生成素对体外培养的大鼠内皮祖细胞功能的影响。方法:提取Wistar大鼠股骨和胫骨骨髓,利用密度梯度离心方法分离单核细胞进行培养,分别在培养基中加入0,4,8U/mL不同浓度的促红细胞生成素进行培养。培养7d后,对内皮祖细胞利用FITC-UEA-1和Dil-Ac-LDL共同染色方法进行鉴定;同时利用CD34和CD133共同标记,以流式细胞仪进行鉴定;采用黏附实验、迁移实验及MTT实验对其功能进行评定。结果与结论:培养到7d左右,细胞形态变化完全,细胞之间形成联接,并可以围成似管腔状形态,十二三天时,细胞融合,形成铺路石样。培养细胞荧光染色显示内皮祖细胞呈双阳性染色,经流式细胞仪鉴定培养的内皮祖细胞可以结合CD133和CD34。黏附实验、迁移实验及MTT实验显示,促红细胞生成素呈剂量依赖性提高内皮祖细胞的黏附、迁移和增殖能力。说明在0～8U/mL浓度内,促红细胞生成素呈剂量依赖性提高内皮祖细胞的黏附、迁移和增殖功能。BACKGROUND：Endothelial progenitor cells（EPCs） is the progenitor cells of endothelial cells,which participated in angiogenesis.Some researches demonstrate that recombinant human erythropoietin（rhEPO） can mobilize EPCs from bone marrow,and have a beneficial effect on recovery of injury tissue.OBJECTIVE：To investigate the effect of rhEPO on the function of rats EPCs.METHODS：Bone marrow was obtained from both femurs and tibials of Wistar rats,and mononuclear cells were isolated by density gradient centrifugation,followed by cultured and treated with several does of rhEPO（0,4,8 U/mL）.At 7 days after culture EPCs were identified by FITC-UEA-1 and Dil-Ac-LDL co-staining.Meanwhile,EPCs were marked by CD133 and CD34,identified by flow cytometry;additionally,the functions of EPCs were evaluated using adhesion test,migration assay and proliferation experiment.RESULTS AND CONCLUSION：At 7 days after culture,the cell morphology changed completely,and formed a tube-like structure,which presented typical cobblestone appearance after cell conjugation.The fluorescence staining showed that the EPCs were positive to both FITC-UEA-1 and Dil-Ac-LDL co-staining.The CD133 and CD34 cells were combined to EPCs by flow cytometry.Finally,the adhesion test,migration assay and proliferation experiment demonstrated that rhEPO could improve adhesion,migration and proliferative capacities in a dose-dependent manner.The results demonstrated that rhEPO can significantly enhance the functions of EPCs in dose-dependent manner at 0-8 U/mL concentration.

关 键 词：内皮祖细胞 促红细胞生成素 细胞培养 功能 大鼠 血管组织工程 

分 类 号：R318[医药卫生—生物医学工程]