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)
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.
收稿日期: 2013-06-05
基金资助:
国家自然科学基金资助项目(81070287); 江苏省自然科学基金资助项目(BK2011484)
作者简介: 党胜春(1972—),男,副教授,副主任医师,博士,主要从事胰腺疾病基础与临床研究。
引用本文:
党胜春1 , 陈纪芳2 ,冯舒2 ,刘彬2 ,王平江2 ,张建新1. 小鼠骨髓源性内皮祖细胞的体外培养及标志物鉴定[J]. 江苏大学学报:医学版, 2013, 23(5): 382-384,388.
DANG Sheng-chun1, CHEN Ji-fang2, FENG Shu2, LIU Bin2, WANG Ping-jiang2, ZHANG Jian-xin1. Culture and identification of mouse bone marrow-derived endothelial progenitor cells in vitro. Journal of Jiangsu University(Medicine Edition), 2013, 23(5): 382-384,388.
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