骨髓源性血管内皮祖细胞的分离培养及鉴定

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。力法：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、VSF，以及摄取DiL-aeLDL的能力进行鉴定。结果：新分离的骨髓单个核细胞呈圆形，2d后细胞贴壁，呈梭形或纺锤形，7d呈集落或线状排列，10d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，并初步建立了骨髓溽性EPCs分离培养、诱导分化及鉴定方法。

Isolation, Culture and Identification of Bone Marrow Derived Endothelial Progenitor Cells

Objective： To explore the rat bone marrow derived endothelial progenitor ceils （EPCs）, culture, differentiation and identification method. Methods： The marrow cavity of rat femur was rinsed, single nuclear ceils was obtained by density gradient centrifugation, those endothelial cells were cultured, the cell morphology was observed. CD34, vWF, CD133 were detected with im- munohistochemistry and immunofluoreseence, absorbed DiL-acLDL and combined with FITC Lectin-UEA-1, uhrastructure and chro- mosome karyotype analysis for identification. Results： The new separation of bone marrow mononuclear cells and rounded. After some cells began to stick wall, a spindle or irregular shape, 7 d show cell colony or linear arrangement, 10 d close to convergence, a typical cobblestone appearance. Adherent cells, CD34, vWF, CD133 positive expression and shows dynamic changes, could absorb Dil acLDL and combined with FITC Lectin UEA-1. Conclusion： This experiment preliminarily established a set of EPCs isolation, culture, differentiation of rat bone marrow.