A high-throughput microfluidic bilayer co-culture platform to study endothelial-pericyte interactions

Miles T Rogers,Lindsay Tomlinson,Robert Gaibler,Joseph L Charest,Matthew P Lech,Rivka Strelnikov,Thomas J Mulhern,Ashley L Gard,Jeffrey T Borenstein,Brian P Cain,Nerses J Haroutunian,Corin Williams,Brett C Isenberg,Nicole E Raustad,Hesham Azizgolshani,Philip M Keegan

doi:10.1038/s41598-021-90833-z

Abstract

Microphysiological organ-on-chip models offer the potential to improve the prediction of drug safety and efficacy through recapitulation of human physiological responses. The importance of including multiple cell types within tissue models has been well documented. However, the study of cell interactions in vitro can be limited by complexity of the tissue model and throughput of current culture systems. Here, we describe the development of a co-culture microvascular model and relevant assays in a high-throughput thermoplastic organ-on-chip platform, PREDICT96. The system consists of 96 arrayed bilayer microfluidic devices containing retinal microvascular endothelial cells and pericytes cultured on opposing sides of a microporous membrane. Compatibility of the PREDICT96 platform with a variety of quantifiable and scalable assays, including macromolecular permeability, image-based screening, Luminex, and qPCR, is demonstrated. In addition, the bilayer design of the devices allows for channel- or cell type-specific readouts, such as cytokine profiles and gene expression. The microvascular model was responsive to perturbations including barrier disruption, inflammatory stimulation, and fluid shear stress, and our results corroborated the improved robustness of co-culture over endothelial mono-cultures. We anticipate the PREDICT96 platform and adapted assays will be suitable for other complex tissues, including applications to disease models and drug discovery.

Highlights

Microphysiological organ-on-chip models offer the potential to improve the prediction of drug safety and efficacy through recapitulation of human physiological responses
Our microvascular model allowed for interaction between the cell types via culturing the Endothelial cells (ECs) and PCs on opposing sides of a 10 μm-thick microporous membrane within the bilayer microfluidic device (Fig. 1C)
To determine the response of the co-culture vascular model to disease or inflammatory perturbations, we performed an inflammation assay using TNFα applied to device channels containing ECs. We studied both “acute” (10 ng/ml TNFα stimulation for 4 h followed by 20 h recovery) and “chronic” dosing (24 h TNFα stimulation), with media collected from each channel at the 24 h time point immediately prior to fixing and staining the samples

Summary

Introduction

Microphysiological organ-on-chip models offer the potential to improve the prediction of drug safety and efficacy through recapitulation of human physiological responses. Progress in the field to date has produced MPS devices that allow for control over the complex biology found in human tissues and organs, such as incorporation of fluid flow to provide media or nutrient replenishment, biomechanical cues such as fluid shear stress (FSS) or stretch to mimic native tissue environments[5,6,7,8,9], and recapitulation of cell-to-fluid ratios in the body to allow for translational readouts[10] High replicate systems, such as the platform described in this work, can facilitate compound screening and high-throughput readouts. We expect the PREDICT96 platform to be advantageous for studying the interactions between multiple cell types in various complex tissue models beyond the microvasculature system described here

Methods

Results

Discussion

Conclusion