A Synthetic Hydrogel, VitroGel ® ORGANOID-3, Improves Immune Cell-Epithelial Interactions in a Tissue Chip Co-Culture Model of Human Gastric Organoids and Dendritic Cells.

Autor:	Cherne MD; Department of Microbiology and Cell Biology, Montana State University, Bozeman, MT, United States., Sidar B; Chemical and Biological Engineering Department and Center for Biofilm Engineering, Montana State University, Bozeman, MT, United States., Sebrell TA; Department of Microbiology and Cell Biology, Montana State University, Bozeman, MT, United States., Sanchez HS; Chemical and Biological Engineering Department and Center for Biofilm Engineering, Montana State University, Bozeman, MT, United States., Heaton K; Department of Microbiology and Cell Biology, Montana State University, Bozeman, MT, United States., Kassama FJ; Department of Chemistry and Biochemistry, Bowdoin College, Brunswick, ME, United States., Roe MM; Department of Microbiology and Cell Biology, Montana State University, Bozeman, MT, United States., Gentry AB; Bozeman GI Clinic, Deaconess Hospital, Bozeman, MT, United States., Chang CB; Chemical and Biological Engineering Department and Center for Biofilm Engineering, Montana State University, Bozeman, MT, United States., Walk ST; Department of Microbiology and Cell Biology, Montana State University, Bozeman, MT, United States., Jutila M; Department of Microbiology and Cell Biology, Montana State University, Bozeman, MT, United States., Wilking JN; Chemical and Biological Engineering Department and Center for Biofilm Engineering, Montana State University, Bozeman, MT, United States., Bimczok D; Department of Microbiology and Cell Biology, Montana State University, Bozeman, MT, United States.
Jazyk:	angličtina
Zdroj:	Frontiers in pharmacology [Front Pharmacol] 2021 Sep 06; Vol. 12, pp. 707891. Date of Electronic Publication: 2021 Sep 06 (Print Publication: 2021).
DOI:	10.3389/fphar.2021.707891
Abstrakt:	Immunosurveillance of the gastrointestinal epithelium by mononuclear phagocytes (MNPs) is essential for maintaining gut health. However, studying the complex interplay between the human gastrointestinal epithelium and MNPs such as dendritic cells (DCs) is difficult, since traditional cell culture systems lack complexity, and animal models may not adequately represent human tissues. Microphysiological systems, or tissue chips, are an attractive alternative for these investigations, because they model functional features of specific tissues or organs using microscale culture platforms that recreate physiological tissue microenvironments. However, successful integration of multiple of tissue types on a tissue chip platform to reproduce physiological cell-cell interactions remains a challenge. We previously developed a tissue chip system, the gut organoid flow chip (GOFlowChip), for long term culture of 3-D pluripotent stem cell-derived human intestinal organoids. Here, we optimized the GOFlowChip platform to build a complex microphysiological immune-cell-epithelial cell co-culture model in order to study DC-epithelial interactions in human stomach. We first tested different tubing materials and chip configurations to optimize DC loading onto the GOFlowChip and demonstrated that DC culture on the GOFlowChip for up to 20 h did not impact DC activation status or viability. However, Transwell chemotaxis assays and live confocal imaging revealed that Matrigel, the extracellular matrix (ECM) material commonly used for organoid culture, prevented DC migration towards the organoids and the establishment of direct MNP-epithelial contacts. Therefore, we next evaluated DC chemotaxis through alternative ECM materials including Matrigel-collagen mixtures and synthetic hydrogels. A polysaccharide-based synthetic hydrogel, VitroGel®-ORGANOID-3 (V-ORG-3), enabled significantly increased DC chemotaxis through the matrix, supported organoid survival and growth, and did not significantly alter DC activation or viability. On the GOFlowChip, DCs that were flowed into the chip migrated rapidly through the V-ORG matrix and reached organoids embedded deep within the chip, with increased interactions between DCs and gastric organoids. The successful integration of DCs and V-ORG-3 embedded gastric organoids into the GOFlowChip platform now permits real-time imaging of MNP-epithelial interactions and other investigations of the complex interplay between gastrointestinal MNPs and epithelial cells in their response to pathogens, candidate drugs and mucosal vaccines. Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. (Copyright © 2021 Cherne, Sidar, Sebrell, Sanchez, Heaton, Kassama, Roe, Gentry, Chang, Walk, Jutila, Wilking and Bimczok.)
Databáze:	MEDLINE
Externí odkaz:	Zobrazit plný text záznamu