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| Culture and identification of mouse bone marrow-derived endothelial progenitor cells in vitro |
| DANG Sheng-chun1, CHEN Ji-fang2, FENG Shu2, LIU Bin2, WANG Ping-jiang2, ZHANG Jian-xin1 |
| （1.Department of General Surgery, the Affiliated Hospital of Jiangsu University, Zhenjiang Jiangsu 212001; 2.Medical School of Jiangsu University, Zhenjiang Jiangsu 212001,China） |
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Abstract [Abstract]Objective: To develope methods of in vitro culture, differentiation and identification of mouse bone marrow derived-endothelial progenitor cells (EPCs)．Methods： Collected EPCs from ICR mouse, differentiated mononuclear cells by density gradient centrifugation，cultured in medium with vascular endothelial growth factor(VEGF), fibroblast growth factor(FGF) and detected the cell surface markers of endothelial cells after 5 days,10 days,15 days,20 days. Results： Cells began to appear epithelial cell morphology after 5 days, and pseudopods appeared;after 10 days of culture, cells began to proliferate, turned round and spindle cells began to appear;after 15 days,cells showed typical morphology of endothelial cells, formed cobblestone like structure，after 20 days,cells performed like long fusiform pulling nets, just similar with mature endothelial cell morphology. Immunofluorescence stain and flow quantitative test results showed, CD34 expression level in EPCs with the extension of incubation time had a gradual weakening trend; VEGFR-2 expression in EPCs with the incubation time had a gradual increasing trend. Conclusions： EPCs from bone marrow cultured in specific conditions could be induced differentiate typical of mature ECs;it was convenient and had strong amplification ability.
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Received: 05 June 2013
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2013, Vol. 23 
