Drug cytotoxicity screening using human intestinal organoids propagated with extensive cost-reduction strategies

• Published in: Scientific reports Vol. 13; no. 1; p. 5407
• Main Authors: Takahashi, Yu, Inoue, Yu, Sato, Shintaro, Okabe, Takayoshi, Kojima, Hirotatsu, Kiyono, Hiroshi, Shimizu, Makoto, Yamauchi, Yoshio, Sato, Ryuichiro
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 03.04.2023; Nature Publishing Group; Nature Portfolio
• Subjects: 631/154; 631/1647; 631/532; 631/61; 631/80; Apoptosis; Caco-2 Cells; Cell culture; Cell Culture Techniques - methods; Cell death; Collagen; Cytotoxicity; Drug development; Embedding; Extracellular signal-regulated kinase; Gene expression; Hepatocyte growth factor; Humanities and Social Sciences; Humans; Intestine; Intestines; Kinases; L cells; MAP kinase; multidisciplinary; Noggin protein; Organoids; Organoids - metabolism; Science; Science (multidisciplinary)
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-023-32438-2

Organoids are regarded as physiologically relevant cell models and useful for compound screening for drug development; however, their applications are currently limited because of the high cost of their culture. We previously succeeded in reducing the cost of human intestinal organoid culture using conditioned medium (CM) of L cells co-expressing Wnt3a, R-spondin1, and Noggin. Here, we further reduced the cost by replacing recombinant hepatocyte growth factor with CM. Moreover, we showed that embedding organoids in collagen gel, a more inexpensive matrix than Matrigel, maintains organoid proliferation and marker gene expression similarly when using Matrigel. The combination of these replacements also enabled the organoid-oriented monolayer cell culture. Furthermore, screening thousands of compounds using organoids expanded with the refined method identified several compounds with more selective cytotoxicity against organoid-derived cells than Caco-2 cells. The mechanism of action of one of these compounds, YC-1, was further elucidated. We showed that YC-1 induces apoptosis through the mitogen-activated protein kinase/extracellular signal-regulated kinase pathway, the mechanism of which was distinct from cell death caused by other hit compounds. Our cost-cutting methodology enables large-scale intestinal organoid culture and subsequent compound screening, which could expand the application of intestinal organoids in various research fields.