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骨髓来源血管内皮前体细胞的培养方法
http://www.100md.com 《第四军医大学学报》 2004年第2期
血管,细胞学,,干细胞;内皮,血管,细胞学;新生血管化;细胞分化,0引言,1材料和方法,2结果,3讨论,【参考文献】
     Focus on cell culture methods of bone marrowderived endothelial progenitor cells

    HAN ShuFang, JIA GuoLiang, WANG HaiYan, ZHANG RongQing

    Department of Cardiovasology, Xijing Hospital, Fourth Military Medical University, Xian 710033, China

    【Abstract】 AIM: To examine whether bone marrow (BM) mononuclear cells (BMMNCs) can give rise to functional EPCs and whether the proliferation and differentation of EPCs are various using different methods. METHODS: BMMNCs were isolated from bonemarrow blood of rat femur by density gradient centrifugation, then divided into 3 groups, plate on culture dishes and maintained EC basal medium: A: Supplement with VEGF/bFGF; B: Coculture with human umbilical vein endothelial cell (HUVEC); C: control group. After 3, 7, 11, 15 d in culture respectively, the cells were used to fluorescent staining and fluorescenceactivated cell sorting to determine the differentiation of the BMMNCs. RESULTS: BMMNCs were isolated and cultured for 7 days resulted in a spindleshaped, EClike morphology. These differentiating cells could be shown to be positive immunostaining for Ⅷ factor relative antigen and NOS3. FACS disclosed that ,the amount of cell and positive immunostaining for factor VIII or NOS3 of B group were significant higher than the other groups (P=0). CONCLUSION: Our findings suggest that BMMNCs isolated from bonemarrow have functional EPCs and can differentiate into ECs in vitro. The adherent cells may contribute to neoangiogenesis in vitro, Bcoculturied with HUVEC can supply cell growth factor at some degree, but can not replace VEGF and bFGF completely. ......

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