摘要
目的 :体外分离纯化人骨髓内皮前体干细胞 ,研究其基本生物学特性 .方法 :利用密度梯度离心法分离成人骨髓得到单个核细胞 ,再用内皮细胞条件培养基培养得到内皮前体干细胞 ,观察细胞生长特性 ,通过检测细胞血管内皮生长因子受体 2 (VEGFR 2 )、Ⅷ因子相关抗原抗体表达和透射电镜对培养细胞进行鉴定 .结果 :原代培养后细胞贴壁生长 ,7～ 10d形成集落或融合呈卵石形 ,免疫组化荧光染色后结果显示VEGFR 2 ,Ⅷ /vWF阳性 ,透射电镜显示细胞内具有特征性的W P小体 .结论 :用密度梯度离心法和条件培养可以从骨髓得到高纯度内皮前体干细胞 ,该细胞具有与血管内皮细胞共同的特征 ,可以进一步分化为血管内皮细胞 ,是理想的组织工程种子细胞 .
AIM: To purify and culture human bone marrow-derived endothelial progenitor cells (EPCs) in vitro and to observe their biological characteristics. METHODS: Mononuclear cells were isolated from bone marrow suspension by density gradient centrifugation and EPCs were harvested in the endothelial growth medium (EGM-2). EPCs were observed under microscope, the cell growth was calculated by cell counting and the cells were identified by electronic microscope examination and immunofluorescence staining for the expression of Ⅷ/vWF, vascular endothelial growth factor receptor-2 (VEGFR-2). RESULTS: The culture-dish-adherent EPCs uniformly had a round or spindle shape and reformed clones with cobble-stone morphology. The cells could be identified by the positive staining for VEGFR-2 and Ⅷ/vWF. Weibel-Paladle bodies could be seen under transmission electron microscope. CONCLUSION: EPCs are one group of stem cells in the bone marrow with the capacity of differentiating into endothelial cells. They can be purified and cultured by density gradient centrifugation and subjected to culture in vitro with endothelial growth medium (EGM-2 MV). These findings suggest that EPCs may be a new seed cell of tissue engineering.
出处
《第四军医大学学报》
北大核心
2004年第20期1834-1837,共4页
Journal of the Fourth Military Medical University
基金
陕西省科技攻关课题 (2 0 0 3K1 0G84)
关键词
骨髓
内皮/细胞学
干细胞
组织工程
种子细胞
bone marrow
endothelium/cytology
stem cells
tissue engineering
seed cells
