Mechanical stimulation and monitoring of artificial collagen membranes for Organ-on Chip applications

Abstract

A novel tool to create artificial, cell-based model systems of biological barriers for preclinical drug testing and basic research are artificial collagen membranes (ACM). They closely reflect in vivo basement membrane properties like permeability and stiffness. Besides, they act as initial artificial basement membrane during cell culture. Nevertheless, up to now only complex protocols are published to create such artificial membranes. Also the deflection relies on very low pressure to elongate ACM, if in vivo like deflection - as in the lung mucosa - shall be emulated. Moreover, online monitoring of the deflection of ACM is a crucial part, hence collagen is non-transparent in the visual spectrum, reducing the applicability of standard microscopy. Here we show an easy protocol to generate ACM using commercially available NetwellTM inserts and collagen gel based coating. To address above mentioned issues in membrane deflection we developed a novel pneumatic actuation device (MPSstimulus) as well as a 3D-printed lid and a holder for 6-well plates as an easy to use microphysiological system (MPS) to stimulate ACM. Furthermore, we characterized the 3D membrane structure and dynamic elongation using optical coherence tomography (OCT). Based on our preliminary results, OCT is a promising technique for high-resolution inspection and elastographic analysis of ACM and adherent cell layers using the novel pneumatic actuation device.

Keywords

• artificial collagen membrane
• optical coherence tomography
• microphysiological system
• biological barrier