Microfluidic co-cultures of retinal pigment epithelial cells and vascular endothelial cells to investigate choroidal angiogenesis

• Published in: Scientific reports 2017-06, Vol.7 (1), p.3538-9, Article 3538
• Main Authors: Chen, Li-Jiun, Ito, Shuntaro, Kai, Hiroyuki, Nagamine, Kuniaki, Nagai, Nobuhiro, Nishizawa, Matsuhiko, Abe, Toshiaki, Kaji, Hirokazu
• Format: Article
• Language: English
• Subjects: 13/106; 13/62; 631/1647/277; 631/61/32; Angiogenesis; Animal models; Cell culture; Cell interactions; Cell Line; Cell Proliferation; Cobalt; Coculture Techniques - instrumentation; Coculture Techniques - methods; Culture Media - chemistry; Endothelial cells; Endothelial Cells - physiology; Epithelial Cells - physiology; Glucose; Glucose - metabolism; Humanities and Social Sciences; Humans; Hypoxia; Lab-On-A-Chip Devices; Macular degeneration; Microfluidics; Monoculture; multidisciplinary; Neovascularization, Pathologic; Oxygen - metabolism; Permeability; Retina; Retinal pigment epithelium; Retinal Pigment Epithelium - physiology; Science; Science (multidisciplinary); Secretion; Umbilical vein; Vascular endothelial growth factor; Vascular Endothelial Growth Factor A - analysis; Zonula occludens-1 protein; Zonula Occludens-1 Protein - analysis
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-017-03788-5

Angiogenesis plays a critical role in many diseases, including macular degeneration. At present, the pathological mechanisms remain unclear while appropriate models dissecting regulation of angiogenic processes are lacking. We propose an in vitro angiogenesis process and test it by examining the co-culture of human retinal pigmental epithelial cells (ARPE-19) and human umbilical vein endothelial cells (HUVEC) inside a microfluidic device. From characterisation of the APRE-19 monoculture, the tight junction protein (ZO-1) was found on the cells cultured in the microfluidic device but changes in the medium conditions did not affect the integrity of monolayers found in the permeability tests. Vascular endothelial growth factor (VEGF) secretion was elevated under low glucose and hypoxia conditions compared to the control. After confirming the angiogenic ability of HUVEC, the cell-cell interactions were analyzed under lowered glucose medium and chemical hypoxia by exposing ARPE-19 cells to cobalt (II) chloride (CoCl 2 ). Heterotypic interactions between ARPE-19 and HUVEC were observed, but proliferation of HUVEC was hindered once the monolayer of ARPE-19 started breaking down. The above characterisations showed that alterations in glucose concentration and/or oxygen level as induced by chemical hypoxia causes elevations in VEGF produced in ARPE-19 which in turn affected directional growth of HUVEC.