Functional intestinal monolayers from organoids derived from human iPS cells for drug discovery research
Abstract Background Human induced pluripotent stem (iPS) cell-derived enterocyte-like cells (ELCs) are expected to be useful for evaluating the intestinal absorption and metabolism of orally administered drugs. However, it is difficult to generate large amounts of ELCs with high quality because they...
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| Format: | Article |
| Language: | English |
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BMC
2024-02-01
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| Series: | Stem Cell Research & Therapy |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13287-024-03685-5 |
| _version_ | 1797275594188128256 |
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| author | Tatsuya Inui Yusei Uraya Jumpei Yokota Tomoki Yamashita Kanae Kawai Kentaro Okada Yukiko Ueyama-Toba Hiroyuki Mizuguchi |
| author_facet | Tatsuya Inui Yusei Uraya Jumpei Yokota Tomoki Yamashita Kanae Kawai Kentaro Okada Yukiko Ueyama-Toba Hiroyuki Mizuguchi |
| author_sort | Tatsuya Inui |
| collection | DOAJ |
| description | Abstract Background Human induced pluripotent stem (iPS) cell-derived enterocyte-like cells (ELCs) are expected to be useful for evaluating the intestinal absorption and metabolism of orally administered drugs. However, it is difficult to generate large amounts of ELCs with high quality because they cannot proliferate and be passaged. Methods To solve the issue above, we have established intestinal organoids from ELCs generated using our protocol. Furthermore, monolayers were produced from the organoids. We evaluated the usefulness of the monolayers by comparing their functions with those of the original ELCs and the organoids. Results We established organoids from ELCs (ELC-org) that could be passaged and maintained for more than a year. When ELC-org were dissociated into single cells and seeded on cell culture inserts (ELC-org-mono), they formed a tight monolayer in 3 days. Both ELC-org and ELC-org-mono were composed exclusively of epithelial cells. Gene expressions of many drug-metabolizing enzymes and drug transporters in ELC-org-mono were enhanced, as compared with those in ELC-org, to a level comparable to those in adult human small intestine. The CYP3A4 activity level in ELC-org-mono was comparable or higher than that in primary cryopreserved human small intestinal cells. ELC-org-mono had the efflux activities of P-gp and BCRP. Importantly, ELC-org-mono maintained high intestinal functions without any negative effects even after long-term culture (for more than a year) or cryopreservation. RNA-seq analysis showed that ELC-org-mono were more mature as intestinal epithelial cells than ELCs or ELC-org. Conclusions We have successfully improved the function and convenience of ELCs by utilizing organoid technology. |
| first_indexed | 2024-03-07T15:16:42Z |
| format | Article |
| id | doaj.art-0d9f9919464a474da5281380fc90759e |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-03-07T15:16:42Z |
| publishDate | 2024-02-01 |
| publisher | BMC |
| record_format | Article |
| series | Stem Cell Research & Therapy |
| spelling | doaj.art-0d9f9919464a474da5281380fc90759e2024-03-05T17:52:12ZengBMCStem Cell Research & Therapy1757-65122024-02-0115111510.1186/s13287-024-03685-5Functional intestinal monolayers from organoids derived from human iPS cells for drug discovery researchTatsuya Inui0Yusei Uraya1Jumpei Yokota2Tomoki Yamashita3Kanae Kawai4Kentaro Okada5Yukiko Ueyama-Toba6Hiroyuki Mizuguchi7Laboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka UniversityLaboratory of Biochemistry and Molecular Biology, School of Pharmaceutical Sciences, Osaka UniversityLaboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka UniversityLaboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka UniversityLaboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka UniversityLaboratory of Biochemistry and Molecular Biology, School of Pharmaceutical Sciences, Osaka UniversityLaboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka UniversityLaboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka UniversityAbstract Background Human induced pluripotent stem (iPS) cell-derived enterocyte-like cells (ELCs) are expected to be useful for evaluating the intestinal absorption and metabolism of orally administered drugs. However, it is difficult to generate large amounts of ELCs with high quality because they cannot proliferate and be passaged. Methods To solve the issue above, we have established intestinal organoids from ELCs generated using our protocol. Furthermore, monolayers were produced from the organoids. We evaluated the usefulness of the monolayers by comparing their functions with those of the original ELCs and the organoids. Results We established organoids from ELCs (ELC-org) that could be passaged and maintained for more than a year. When ELC-org were dissociated into single cells and seeded on cell culture inserts (ELC-org-mono), they formed a tight monolayer in 3 days. Both ELC-org and ELC-org-mono were composed exclusively of epithelial cells. Gene expressions of many drug-metabolizing enzymes and drug transporters in ELC-org-mono were enhanced, as compared with those in ELC-org, to a level comparable to those in adult human small intestine. The CYP3A4 activity level in ELC-org-mono was comparable or higher than that in primary cryopreserved human small intestinal cells. ELC-org-mono had the efflux activities of P-gp and BCRP. Importantly, ELC-org-mono maintained high intestinal functions without any negative effects even after long-term culture (for more than a year) or cryopreservation. RNA-seq analysis showed that ELC-org-mono were more mature as intestinal epithelial cells than ELCs or ELC-org. Conclusions We have successfully improved the function and convenience of ELCs by utilizing organoid technology.https://doi.org/10.1186/s13287-024-03685-5Intestinal organoidsSmall intestineiPS cellDrug-metabolizing enzymesDrug transporters |
| spellingShingle | Tatsuya Inui Yusei Uraya Jumpei Yokota Tomoki Yamashita Kanae Kawai Kentaro Okada Yukiko Ueyama-Toba Hiroyuki Mizuguchi Functional intestinal monolayers from organoids derived from human iPS cells for drug discovery research Stem Cell Research & Therapy Intestinal organoids Small intestine iPS cell Drug-metabolizing enzymes Drug transporters |
| title | Functional intestinal monolayers from organoids derived from human iPS cells for drug discovery research |
| title_full | Functional intestinal monolayers from organoids derived from human iPS cells for drug discovery research |
| title_fullStr | Functional intestinal monolayers from organoids derived from human iPS cells for drug discovery research |
| title_full_unstemmed | Functional intestinal monolayers from organoids derived from human iPS cells for drug discovery research |
| title_short | Functional intestinal monolayers from organoids derived from human iPS cells for drug discovery research |
| title_sort | functional intestinal monolayers from organoids derived from human ips cells for drug discovery research |
| topic | Intestinal organoids Small intestine iPS cell Drug-metabolizing enzymes Drug transporters |
| url | https://doi.org/10.1186/s13287-024-03685-5 |
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