Enhanced Small Intestinal Organoid-derived Epithelial Cell Adhesion and Growth in Organ-on-a-chip Devices

Organ-on-a-chip devices are predominately made of the polymer polymethylsiloxane (PDMS), exhibiting several attractive properties , transparency, gas permeability, and biocompatibility. However, the attachment of cells to this polymer has proven challenging, especially for delicate primary cells , small intestinal organoid-derived epithelial cells. Hence, a need to functionalize and coat the surface has arisen to render it more hydrophilic and improve its ability to support cell adhesion and growth. While previous research has demonstrated some successful results in culturing primary cells, no comprehensive and comparative protocol has been proposed. Here, we provide a protocol for enhanced small intestinal organoid-derived epithelial cell adhesion and growth on PDMS and plastics, assessing both PDMS surface functionalization, adhesion protein coating as well as medium selection. We assess PDMS functionalization using (3-aminopropyl)trimethoxysilane (APTMS) or polyethyleneimine-glutaraldehyde (PEIGA), and adhesion protein coating using various Laminins, Collagen I, Matrigel, or mixtures thereof. Finally, we assess the use of two different medium compositions including growth factors EGF, Noggin and R-spondin1 (ENR medium) alone or combined with the two small molecules CHIR99021 and valproic acid (CV medium). We envision that our results will be useful for further attempts in emulating the small intestine using plastic- or PDMS-based devices for organs-on-a-chip development.