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目的:探索大鼠骨髓源性血管内皮祖细胞(endothelial progenitor cells,EPCs)的培养、诱导分化及鉴定方法。方法:取大鼠股骨并冲洗骨髓腔,梯度密度离心法获单个核细胞后内皮细胞培养液培养,通过细胞形态、免疫组化和免疫荧光检测CD34、vWF、CD133、ⅧF,以及摄取DiL-acLDL的能力进行鉴定。结果:新分离的骨髓单个核细胞呈圆形,2 d后细胞贴壁,呈梭形或纺锤形,7 d呈集落或线状排列,10 d接近融合,呈鹅卵石样排列;贴壁细胞CD34、ⅧF、vWF、CD133均表达阳性,能摄取Dil-acLDL并结合FITC-Lectin-UEA-1。结论:从大鼠骨髓中分离出EPCs,并初步建立了骨髓源性EPCs分离培养、诱导分化及鉴定方法。
Objective: To explore the culture, differentiation and identification of rat bone marrow-derived endothelial progenitor cells (EPCs). Methods: The femoral bone of rabbits was flushed and the bone marrow cavity was washed. The culture medium of endothelial cells was obtained by gradient density centrifugation after mononuclear cells were cultured. CD34, vWF, CD133 and ⅧF were detected by cell morphology, immunohistochemistry and immunofluorescence staining and DiL-acLDL The ability to identify. Results: The newly isolated bone marrow mononuclear cells were round in shape. After 2 days, the cells adherent, spindle-shaped or spindle-shaped, arranged in a colony or line shape on the 7th day and arranged in a cobblestone pattern on the 10th day. The adherent cells CD34 , ⅧF, vWF and CD133, all of which could uptake Dil-acLDL and bind to FITC-Lectin-UEA-1. CONCLUSION: EPCs were isolated from bone marrow of rats and the methods of isolation, culture, differentiation and identification of bone marrow-derived EPCs were initially established.