Korean J Ophthalmol.  2018 Feb;32(1):70-76. 10.3341/kjo.2016.0115.

Relationship between Pericytes and Endothelial Cells in Retinal Neovascularization: A Histological and Immunofluorescent Study of Retinal Angiogenesis

  • 1Department of Ophthalmology, Seoul National University Hospital, Seoul National University College of Medicine, Seoul, Korea. ysyu@snu.ac.kr
  • 2Mechanical Engineering, Seoul National University, Seoul, Korea.
  • 3Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul, Korea.
  • 4FARB (Fight against Angiogenesis-Related Blindness) Laboratory, Clinical Research Institute, Seoul National University Hospital, Seoul, Korea.


To evaluate the relationship between pericytes and endothelial cells in retinal neovascularization through histological and immunofluorescent studies.
C57BL/6J mice were exposed to hyperoxia from postnatal day (P) 7 to P12 and were returned to room air at P12 to induce a model of oxygen-induced retinopathy (OIR). The cross sections of enucleated eyes were processed with hematoxylin and eosin. Immunofluorescent staining of pericytes, endothelial cells, and N-cadherin was performed. Microfluidic devices were fabricated out of polydimethylsiloxane using soft lithography and replica molding. Human retinal microvascular endothelial cells, human brain microvascular endothelial cells, human umbilical vein endothelial cells and human placenta pericyte were mixed and co-cultured.
Unlike the three-layered vascular plexus found in retinal angiogenesis of a normal mouse, angiogenesis in the OIR model is identified by the neovascular tuft extending into the vitreous. Neovascular tufts and the three-layered vascular plexus were both covered with pericytes in the OIR model. In this pathologic vascularization, N-cadherin, known to be crucial intercellular adhesion molecule, was also present. Further evaluation using the microfluidic in vitro model, successfully developed a microvascular network of endothelial cells covered with pericytes, mimicking normal retinal angiogenesis within 6 days.
Pericytes covering endothelial cells were observed not only in vasculature of normal retina but also pathologic neovascularization of OIR mouse at P17. Factors involved in the endothelial cell-pericyte interaction can be evaluated as an attractive novel treatment target. These future studies can be performed using microfluidic systems, which can shorten the study time and provide three-dimensional structural evaluation.


Endothelial cells; Microfluidics; Oxygen induced retinopathy; Pericytes; Retinal neovascularization
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