【摘要】 目的建立分离小鼠骨髓内皮祖细胞(endothelial progenitor cell,EPC)的方法。方法通过密度梯度分离法从小鼠骨髓中分离单个核细胞,行流式细胞检测,培养7d后,行DIL-acLDL和FITC-UEA-1双荧光染色、细胞移行实验和血管内皮网络掺入实验。结果流式细胞技术检测结果显示,骨髓来源的CD34/CD133/VEGFR2三阳细胞为0.029%±0.008%;细胞培养7d后,可见梭形细胞呈优势生长;DIL-acLDL与FITC-UEA-1荧光标记的双阳性细胞占细胞总数的比例为86.085%±5.622%;细胞移行实验结果可见平均每视野移行细胞数为(19.458±2.251)个;血管内皮网络掺入实验结果为,每视野掺入细胞数平均为(67.750±8.823)个。结论用流式细胞技术检测分离小鼠骨髓单个核细胞不同表面标志物的表达,用选择性内皮生长体系培养EPC,再通过细胞免疫及细胞功能检测所分离的细胞,是一种较为理想的分离与鉴定EPC的方法。更多还原

【Abstract】 Objective To investigate an appropriate method for isolating EPCs from mice bone marrow.Methods The mononuclear cells from mice bone marrow collected by density gradient separation were cultured in M199 medium after examination by flow cytometry.After 7 days’ culture,cells were stained with DIL-acLDL and FITC-UEA-1,and tested the abilities of migration and incorporation into the vascular network.Results The spindle cells grew dominantly after 7 days’ culture.About 0.029%±0.008% of total mononuclear cells detected by flow cytometry were CD34+CD133+VEGFR2+ cells.The percentage of cells labeled both with DIL-acLDL and FITC-UEA-1 was 86.085%±5.622%.The number of migratory cells reached 19.458±2.251/field of view.The average cell number incorporating into the vascular network was 67.750±8.823/field of view detected by cell migration test.Conclusion The above method is a relatively ideal one for isolating and assessing EPCs from mice bone marrow,which could offer a better support to further research on EPCs.更多还原