Establishment of three-dimensional model of rat retinal angiogenesis in vitro based on endothelial cells and pericytes

Abstract

AIM: To establish three-dimensional(3D)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 12h of co-culture, RMPs were recruited by ECs and gathered into cell masses with different sizes. At 24h, ECs/RMPs formed a complex 3D vascular spline network. At 48h, the reticular structure disintegrated obviously, and only a small amount of incomplete and simple reticular structure remained. At 72h, the vascular spline cable network disintegrated completely. In the development of 3D model, the expression of VEGF-A increased, but decreased when it degenerated. CONCLUSION: This study successfully established a 3D model of rat retinal angiogenesis in vitro based on ECs and RMPs.

Keywords

• retinal microvessels
• endothelial cells
• pericytes
• angiogenesis
• three-dimensional model