Table 3.
Overview of the currently established organoid–cell co-culture systems.
| Intestinal cells | Co-culture | Device material | Technique | Major observations | Ref. |
|---|---|---|---|---|---|
| Caco-2 | L. rhamnosus | PDMS, type I collagen, and Matrigel mix | Soft lithography | Fluid flow accelerated intestinal epithelial differentiation and organization into villi-like structures, mechanical stimulation enhanced specific differentiation features, sustained long-term co-culture with commensal bacteria | [196] |
| Human duodenal organoids (pediatric donors) | HIMECs | PDMS, type I collagen, and Matrigel mix | Soft lithography | Transcriptome of the intestinal tissue-on-chip more closely resembled that of the duodenum in vivo than the initial organoid culture from which it was derived, co-culture with endothelial cells accelerated the formation of the epithelial monolayer | [184] |
| Human duodenal organoids (adult donors) | HIMECs | PDMS, type IV collagen, and Matrigel mix | Soft lithography | Showed culture system suitability for studying intestinal metabolism and drug transport | [197] |
| Human jejunal organoids (adult donors) | HUVECs | PDMS, type IV collagen | Soft lithography | Shear stress generated by luminal and basolateral flow produced a model of continuous intestinal differentiation, no villi-like structures observed with stem cell expansion media on the luminal side | [198] |
| Mouse colon tissue explant | Intestinal submucosal and muscular layers, microbiota |
Cyclin olefin polymer and polyurethane no ECM |
Injection molding | Dual-flow microfluidics allowed for the culture of full thickness explants over 3 days, recapitulated the in vivo oxygen gradient across the epithelial layer | [199] |
| Mouse proximal small intestine organoids | Cryptosporidium parvum | PDMS, Type I collagen, and Matrigel mix-coated 3D scaffold | Soft lithography and laser ablation | Established a long-lived and tube-shaped intestinal epithelial culture system by using crypt-like microcavities under flow, induced topography-guided self-organization of a functional epithelium with crypt- and villus-like domains similar to that observed in vivo, the culture system showed self-regeneration capacity and response to bacterial infection | [200] |
| Caco-2 | L. rhamnosus GG and Bacteroides caccae | Polycarbonate, type I collagen; porcine gastric mucin | Computer-controlled milling, laser cutting, and bolting | Engineered a modular architecture consisting of 3 microchambers to facilitate human and microbial cell interface, allowed measuring individual transcriptional responses in different infectious contexts and real-time monitoring of oxygen concentrations | [201] |
| Caco-2 | Human gut microbiota, E. coli, human PBMCs, human microvascular endothelial cells, and human lymphatic microvascular endothelial cells | PDMS, type I collagen, and Matrigel mix | Soft lithography | Established a stable long-term co-culture system of commensal and pathogenic microbes with intestinal epithelial cells, lack of mechanical stimulation induced bacterial overgrowth, similar to what is observed in IBD patients, emulated intestinal infection and inflammatory responses | [180] |
| Caco-2 | Human gut microbiota, E. coli, PBMCs | PDMS, type I collagen, and Matrigel mix | Soft lithography | Re-created a dextran sodium sulfate–induced epithelial inflammatory response, described intestinal barrier dysfunction as a critical trigger of inflammation onset in the gut | [202] |
| Caco-2 | S. flexneri | PDMS, type I collagen, and Matrigel mix | Soft lithography | Enabled the replication of Shigella infection hallmarks, Shigella invaded directly via the luminal side of the epithelium composed solely of enterocytes, 3D crypt-like structures provided a safe harbor for bacteria against luminal washout | [203] |
| Human colon organoids (pediatric and adult donors) | HIMECs, EHEC | PDMS, type I collagen, and Matrigel mix | Soft lithography | Observed that infectious activity of EHEC is promoted by human gut microbiome metabolites, when compared with those derived from mouse, recapitulated the proinflammatory and anti-inflammatory cytokine profiles induced by EHEC infection | [204] |
| Caco-2 | Bifidobacterium adolescentis and Eubacterium hallii | PDMS, type I collagen and Matrigel mix | Soft lithography | Simulated a steady-state vertical oxygen gradient, the transepithelial anoxic interface allowed co-culture with obligate anaerobes | [205] |
| Caco-2 and human ileal organoids (pediatric donors) | Bacteroides fragilis, human gut microbiota, HIMECs | PDMS, type I collagen, and Matrigel mix | Soft lithography | Established an oxygen gradient compatible with co-culture of a complex community of anaerobic commensal microorganisms | [183] |
| Caco-2 | HUVECs, PBMCs, mucosal macrophages, dendritic cells, L. rhamnosus, Candida albicans | Polystyrol, PET | Injection molding | Characterized immunologic responses to luminal lipopolysaccharide and endotoxemia, addressed the role of probiotics in protecting from opportunistic infections | [206] |
| Caco-2 | Neutrophils, monocytic THP1 cells |
Glass, polystyrene, and proprietary polymers Membrane-free (Phase Guide) type I collagen |
— | Simulated acute intestinal inflammatory responses by enabling neutrophil recruitment to the parenchymal compartment | [38] |
| Human colon organoids (pediatric and adult donors) | PBMCs |
Glass, polystyrene, and proprietary polymers type I collagen |
— | Studied IBD-associated inflammatory responses | [207] |
| Human ileal organoids (pediatric donors) | Endothelial colony forming cell derived endothelial cells, intestinal subepithelial myofibroblasts | PDMS, Polycarbonate type IV collagen | Soft lithography | Characterized ISEMF-induced angiogenesis and its physiological parameters, evaluated the effect of perfused vasculature on intestinal epithelial cell culture | [208] |
| Mouse small intestine | Lamina propria lymphocytes and ILC3s | recombinant mouse IL-2, IL-7, IL-15 and IL-23 | — | IL-22-dependent stem cell proliferation | [123] |
| Mouse small intestine | Pre-stimulated CD4+ splenocytes | Organoid medium | — | Intestinal stem cell differentiation | [119] |
| Human fetal intestine | Pre-stimulated fetal lamina propria T cells | p38 MAPK inhibitor, recombinant human IL-2 | — | Organoid outgrowth | [209] |
| Mouse small intestine | αβ and γδ IELs | recombinant mouse IL-2, IL-7, IL-15 | — | IEL survival, proliferation, and incorporation in the epithelium | [210] |
| Mouse small intestine | OT-I CD8+ splenocytes | EGF, noggin, R-spondin1 or R-spondin3 | — | T cell proliferation and IEL phenotype | [211] |
| Mouse Lgr5(+) ISCs at single-cell level | OT-II splenocytes (naive) | Organoid medium | — | T cell proliferation | [119] |
CAR chimeric antigen receptor, CDK cyclin-dependent kinase, CRC colorectal cancer, DC dendritic cell, DKK1 dickkopf Wnt signaling pathway inhibitor 1, ECM extracellular matrix, EGF epidermal growth factor, EHEC enterohemorrhagic E coli, FZD9 frizzled class receptor 9, HIMEC human intestinal microvascular endothelial cell, HUVEC human umbilical vein endothelial cell, IEL intraepithelial lymphocyte, ILC3 group 3 innate lymphoid cell, ISEMF intestinal subepithelial myofibroblast, MAPK mitogen-activated protein kinase, MDOTS mouse-derived organotypic tumor spheroid, MUC2 mucin 2, NSCLC non-small-cell lung cancer, PD1 programmed cell death 1, PDMS polydimethylsiloxane, PDOTS patient-derived organotypic tumor spheroid, PET polyethylene terephthalate, RSV respiratory syncytial virus, TCR T cell receptor, THP1 Tohoku Hospital Pediatrics-1 cell line, WNT3A WNT family member 3A.