血管内皮生长因子诱导的胶质瘤源性微血管内皮细胞体外三维成型特性

VEGF-induced tubulogenesis of endothelial cells from human brain malignant glioma in the three dimentional model

目的比较人脑胶质瘤源性微血管内皮细胞(GDMEC)和内皮细胞样细胞ECV304三维培养血管生成特性,探讨GDMEC在血管生成研究中的意义。方法采用免疫磁珠分选系统获得纯化的人脑GDMEC,以胶原为介质建立内皮细胞三维培养模型,比较观察GDMEC与ECV304细胞三维培养小管样结构(TLS)形成及不同浓度血管内皮生长因子(VEGF)对两种细胞TLS形成的诱导作用。结果所获GDMEC细胞纯度达98%,可连续传代培养。GDMEC的TLS形成数显著多于同数量、同时相点ECV304细胞的TLS形成数量。VEGF在体外对GDMEC有显著的促进TLS形成作用,且呈量效和时效关系;VEGF对ECV304细胞形成小管样结构的诱导作用弱。结论GDMEC在体外保持了内皮细胞特征和活跃的血管生成特性,对VEGF的反应性较ECV304好,因而GDMEC更适合体外血管生成研究。

Objective To compare the tubulogenesis capability of malignant glioma-derived microvessel endothelial cells （GDMEC） from human brain with that of ECV304 cells in a three dimentional model and to explore the significance of GDMEC in the study on angiogenesis. Methods The GDMEC were isolated from malignant gliomas of human brain and purified by selectS, re binding to the monoclonal antibody against CD105 bound to the magnetic MACS MicroBeads. GDMEC and endothelial-like cell line ECV304 were compared with their capabilities of formatting tubule-like structure （TLS） in the three dimentional collagen matrix, with or without inducement by various concentration of vascular endothelial growth factor （VEGF）. Results The obtained GDMEC had a high purification （98%） and could be successfully cultured in vitro. GDMECs formed more TLS than ECV304 cells of the same number and at the same time points. VEGF could induce rapid formation of TLS in a dose-dependent manner, however, ECV304 cells were less response to VEGF stimulation. Conclusions GDMEC could maintain their endothelial characteristics and potential capability of angiogenesis. They were more response to VEGF than ECV304, therefore, more suitable for in vitro studies on tumor angiogenesis.