【摘要】 目的:构建基于视网膜微血管内皮细胞(ECs)和周细胞(RMPs)的大鼠视网膜血管新生体外三维模型。方法:分离、纯化、培养大鼠视网膜微血管内皮细胞及视网膜微血管周细胞,选用第3～7代经鉴定后的细胞用于研究,采用细胞示踪剂对细胞进行染色,混合后按表面培养法接种于Matrigel,动态观察,并检测血管新生期间血管内皮生长因子A(VEGF-A)的表达。结果:培养12h时,RMPs被ECs招募,聚集成大小不等细胞团块;24h时,ECs/RMPs共同形成复杂的三维血管样条索网;48h时,网状结构出现明显崩解,仅保持少量不完整的简单网状结构;72h时,血管样条索网完全崩解。在三维模型发生过程中,VEGF-A的表达量升高;退化时,VEGF-A的表达量下降。结论:成功基于视网膜微血管全成分细胞ECs和RMPs构建了大鼠视网膜血管新生体外三维模型。更多还原

【Abstract】 AIM:To establish three-dimensional(3 D) model of rat retinal angiogenesis in vitro based on retinal microvascular endothelial cells(ECs) and retinal microvascular pericytes(RMPs).METHODS:The identified ECs and RMPs of third generation to seventh generation were used for research after isolated,purified and cultured.The cells were stained with cell tracer.Then,it were mixed and inoculated on Matrigel by the surface culture method for dynamic observation.The expression of VEGF-A was assessed during angiogenesis.RESULTS:At 12 h of co-culture,RMPs were recruited by ECs and gathered into cell masses with different sizes.At 24 h,ECs/RMPs formed a complex 3 D vascular spline network.At 48 h,the reticular structure disintegrated obviously,and only a small amount of incomplete and simple reticular structure remained.At 72 h,the vascular spline cable network disintegrated completely.In the development of 3 D model,the expression of VEGF-A increased,but decreased when it degenerated.CONCLUSION:This study successfully established a 3 D model of rat retinal angiogenesis in vitro based on ECs and RMPs.更多还原