Abstract
Microfluidic-based organ-on-a-chip assays with simultaneous coculture of multi-cell types have been widely utilized for basic research and drug development. Here we describe a novel method for a primary cell-based corneal microphysiological system which aims to recapitulate the basic functions of the in vivo cornea and to study topically applied ocular drug permeation. In this study, the protocols for isolating and cultivating primary corneal epithelial cells and endothelial cells from mouse inbred strain C57BL/6J were optimized, to allow for the development of a primary-cell based microfluidic 3D micro-engineered cornea. This tissue unit, by overcoming the limitations of 2D conventional cell culture, supports new investigations on cornea function and facilitates drug delivery testing.
Highlights
The cornea is an avascular and transparent mucosal tissue, and serves as one of the body's major mechanical barriers (DelMonte and Kim, 2011; Sridhar, 2018)
Corneal Endothelial Cells One day prior to seeding endothelial cells into microfluidic devices, the peripheral microfluidic channel was coated with collagen IV solution for at least 1h in 37°C incubator
We have improved the current corneal epithelial cell culture protocol by an integration of tissue explant and enzyme digestion methods and accommodated these cells to grow in microfluidic chambers
Summary
The cornea is an avascular and transparent mucosal tissue, and serves as one of the body's major mechanical barriers (DelMonte and Kim, 2011; Sridhar, 2018). To model the in vivo system, a condensed collagen layer was formed in the epithelium channel to mimic Bowman's layer with the concept of viscous finger (Bischel et al, 2012), a method to produce hydrogel lumen structure (Chin et al, 2002) This design is highly accessible to most of the standard biological labs and would provide a precise model to study physiological/pathological conditions of cornea tissues for ophthalmological drug discovery, potentially leading to development of novel ocular drug delivery methods across the anterior chamber. Cell TrackerTM Red (Thermo Fisher Scientific, Cat. No.C34552).
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