Abstract
Organoids, a multi-cellular and organ-like structure cultured in vitro, can be used in a variety of fields such as disease modeling, drug discovery, or cell therapy development. When organoids derived from Lgr5 stem cells are cultured ex vivo, recombinant R-spondin-1 protein should be added at a high concentration for the initiation and maintenance of the organoids. Because the addition of large amounts of R-spondin-1 greatly increases the cost of organoids, the organoids grown with R-spondin-1 are not practical for large-scale drug screening and for the development of therapeutic agents. In this study, we tried to find a R-spondin-1 substitute compound that is able initiate small intestinal organoids without the use of the R-spondin-1 protein; thus, using organoid media that each included one compound from among an 8,364 compound library instead of R-spondin-1, we observed whether organoids were established from the crypts of the small intestine. As a result, we found one compound that could promote the initial formation and growth of enteroids in the medium without R-spondin-1 and named it RS-246204. The enteroids grown with RS-246204 had a similar differentiation capacity as well as self-renewal capacity as the enteroids grown with R-spondin-1. Furthermore, the RS-246204-derived enteroids could successfully produce the forskolin induced swelling and the organoid based epithelial to mesenchymal transition model. This compound could be used for developing a cost-efficient culturing method for intestinal organoids as well as for exploring Lgr5 signaling, intestinal stem cell physiology and therapeutics for GI tract diseases.
Highlights
An organoid is a group of cells cultured in vitro, which has a cell population, structure and organ specific function similar to organs [1]
The enteroids cultured with EN-RS246204 had a similar growth (Figure 2D–2E), spatial organization (Figure 3) and intestinal specific function (Figures 5, 6) as the intestinal enteroids cultured with ENR
The R-spondin / leucine-rich repeat-containing G-proteincoupled receptor5 (Lgr5) / β-catenin module is known to be a positive regulator of the Wnt signal strength, which is involved in the self-renewal of intestinal stem cells and maintenance of tissue homeostasis [25]
Summary
An organoid is a group of cells cultured in vitro, which has a cell population, structure and organ specific function similar to organs [1]. It is possible to model a variety of diseases producing intestinal fibrosis models by treating the organoid with several cytokines [4] as well as colorectal cancer models by introducing cancer related mutations via CRISPR/Cas9 [5] and inflammatory bowel disease (IBD) model using samples derived from patients [6, 7]. These models can provide www.impactjournals.com/oncotarget a good platform for studying pathophysiology, screening drug efficacy, and testing drug toxicity. The organoid may be used to develop therapeutic agents to regenerate damaged tissue
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