A Microwell-Based Intestinal Organoid-Macrophage Co-Culture System to Study Intestinal Inflammation

Abstract

The mammalian intestinal epithelium contains more immune cells than any other tissue, and this is largely because of its constant exposure to pathogens. Macrophages are crucial for maintaining intestinal homeostasis, but they also play a central role in chronic pathologies of the digestive system. We developed a versatile microwell-based intestinal organoid-macrophage co-culture system that enables us to recapitulate features of intestinal inflammation. This microwell-based platform facilitates the controlled positioning of cells in different configurations, continuous in situ monitoring of cell interactions, and high-throughput downstream applications. Using this novel system, we compared the inflammatory response when intestinal organoids were co-cultured with macrophages versus when intestinal organoids were treated with the pro-inflammatory cytokine TNF-α. Furthermore, we demonstrated that the tissue-specific response differs according to the physical distance between the organoids and the macrophages and that the intestinal organoids show an immunomodulatory competence. Our novel microwell-based intestinal organoid model incorporating acellular and cellular components of the immune system can pave the way to unravel unknown mechanisms related to intestinal homeostasis and disorders.

Attenuation of Chronic Inflammation in Intestinal Organoids with Graphene Oxide-Mediated Tumor Necrosis Factor-α_Small Interfering RNA Delivery

“…This observation aligns with previous research indicating that treatment of organoids with 64 ng/mL of TNF-α induced the production of TNF-α and associated inflammatory cytokines. 70 In summary, these findings collectively establish the successful induction of a robust inflammatory phenotype closely resembling that observed in IBD. This model serves as a valuable tool for investigating the mechanisms underlying chronic inflammation and evaluating potential therapeutic interventions.…”

“…Importantly, the inflammatory cocktail induced increased secretion levels of TNF-α, IL-6, and IL-1β compared with the untreated controls. This observation aligns with previous research indicating that treatment of organoids with 64 ng/mL of TNF-α induced the production of TNF-α and associated inflammatory cytokines . In summary, these findings collectively establish the successful induction of a robust inflammatory phenotype closely resembling that observed in IBD.…”

A Modular Microfluidic Organoid Platform Using LEGO‐Like Bricks

“…The polymeric carrier is a PC membrane (50 μm in thickness) that can be microthermoformed to include different patterns, such as trays, pockets, or microcavities. [30,31] Moreover, multiple cell types can be directly cultured onto the PC carrier [32] and on both sides. [33] For example, this offers the opportunity to create a vascular barrier beneath the tissue model off-chip, which once matured, can be transferred inside the OoC device.…”

Human Pluripotent Stem Cell-Derived Alveolar Organoids: Cellular Heterogeneity and Maturity

“…In the case of brain organoids, studies on immune cell-containing organoid systems were successfully conducted through direct co-culture with microglia [ 51 , 52 ]. In addition, intestinal organoids have been actively applied to homeostasis regulation and disease mechanism studies by generating 3D multicellular organoids [ 53 , 54 ]. However, there are still challenges to be resolved to recapitulate the cellular diversity of human tissue, and there have been few reports on the function, maintenance, and disease modeling of organoids, including their populations of immune cells and other cells, especially in pulmonary AOs.…”