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Generation of human induced pluripotent stem cell-derived liver buds with chemically defined and animal origin-free media

Advances in organoid technology have broadened the number of target diseases and conditions in which human induced pluripotent stem cell (iPSC)-based regenerative medicine can be applied; however, mass production of organoids and the development of chemically defined, animal origin-free (CD-AOF) med...

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Autores principales: Sekine, Keisuke, Ogawa, Shimpei, Tsuzuki, Syusaku, Kobayashi, Tatsuya, Ikeda, Kazuki, Nakanishi, Noriko, Takeuchi, Kenta, Kanai, Eriko, Otake, Yugo, Okamoto, Satoshi, Kobayashi, Tsuyoshi, Takebe, Takanori, Taniguchi, Hideki
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7578079/
https://www.ncbi.nlm.nih.gov/pubmed/33087763
http://dx.doi.org/10.1038/s41598-020-73908-1
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author Sekine, Keisuke
Ogawa, Shimpei
Tsuzuki, Syusaku
Kobayashi, Tatsuya
Ikeda, Kazuki
Nakanishi, Noriko
Takeuchi, Kenta
Kanai, Eriko
Otake, Yugo
Okamoto, Satoshi
Kobayashi, Tsuyoshi
Takebe, Takanori
Taniguchi, Hideki
author_facet Sekine, Keisuke
Ogawa, Shimpei
Tsuzuki, Syusaku
Kobayashi, Tatsuya
Ikeda, Kazuki
Nakanishi, Noriko
Takeuchi, Kenta
Kanai, Eriko
Otake, Yugo
Okamoto, Satoshi
Kobayashi, Tsuyoshi
Takebe, Takanori
Taniguchi, Hideki
author_sort Sekine, Keisuke
collection PubMed
description Advances in organoid technology have broadened the number of target diseases and conditions in which human induced pluripotent stem cell (iPSC)-based regenerative medicine can be applied; however, mass production of organoids and the development of chemically defined, animal origin-free (CD-AOF) media and supplements are unresolved issues that hamper the clinical applicability of these approaches. CD-AOF media and supplements ensure the quality and reproducibility of culture systems by lowering lot-to-lot variations and the risk of contamination with viruses or toxins. We previously generated liver organoids from iPSCs, namely iPSC-liver buds (iPSC-LBs), by mimicking the organogenic interactions among hepatocytes, endothelial cells (ECs), and mesenchymal cells (MCs) and recently reported the mass production of iPSC-LBs derived entirely from iPSCs (all iPSC-LBs), which should facilitate their large-scale production for the treatment of liver failure. However, in previous studies we used media originating from animals for differentiation except for the maintenance of undifferentiated iPSCs. Therefore, we developed a CD-AOF medium to generate all iPSC-LBs. We first developed a CD-AOF medium for hepatocytes, ECs, and stage-matched MCs, i.e., septum transversum mesenchyme (STM), in 2D cultures. We next generated all iPSC-LBs by incubating individual cell types in ultra-low attachment micro-dimple plates. The hepatic functions of all iPSC-LBs generated using the CD-AOF medium were equivalent to those of all iPSC-LBs generated using the conventional medium both in vitro and in vivo. Furthermore, we found that this CD-AOF medium could be used in several cell culture settings. Taken together, these results demonstrate the successful development of a CD-AOF medium suitable for all iPSC-LBs. The protocol developed in this study will facilitate the clinical applicability of all iPSC-LBs in the treatment of liver diseases.
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spelling pubmed-75780792020-10-23 Generation of human induced pluripotent stem cell-derived liver buds with chemically defined and animal origin-free media Sekine, Keisuke Ogawa, Shimpei Tsuzuki, Syusaku Kobayashi, Tatsuya Ikeda, Kazuki Nakanishi, Noriko Takeuchi, Kenta Kanai, Eriko Otake, Yugo Okamoto, Satoshi Kobayashi, Tsuyoshi Takebe, Takanori Taniguchi, Hideki Sci Rep Article Advances in organoid technology have broadened the number of target diseases and conditions in which human induced pluripotent stem cell (iPSC)-based regenerative medicine can be applied; however, mass production of organoids and the development of chemically defined, animal origin-free (CD-AOF) media and supplements are unresolved issues that hamper the clinical applicability of these approaches. CD-AOF media and supplements ensure the quality and reproducibility of culture systems by lowering lot-to-lot variations and the risk of contamination with viruses or toxins. We previously generated liver organoids from iPSCs, namely iPSC-liver buds (iPSC-LBs), by mimicking the organogenic interactions among hepatocytes, endothelial cells (ECs), and mesenchymal cells (MCs) and recently reported the mass production of iPSC-LBs derived entirely from iPSCs (all iPSC-LBs), which should facilitate their large-scale production for the treatment of liver failure. However, in previous studies we used media originating from animals for differentiation except for the maintenance of undifferentiated iPSCs. Therefore, we developed a CD-AOF medium to generate all iPSC-LBs. We first developed a CD-AOF medium for hepatocytes, ECs, and stage-matched MCs, i.e., septum transversum mesenchyme (STM), in 2D cultures. We next generated all iPSC-LBs by incubating individual cell types in ultra-low attachment micro-dimple plates. The hepatic functions of all iPSC-LBs generated using the CD-AOF medium were equivalent to those of all iPSC-LBs generated using the conventional medium both in vitro and in vivo. Furthermore, we found that this CD-AOF medium could be used in several cell culture settings. Taken together, these results demonstrate the successful development of a CD-AOF medium suitable for all iPSC-LBs. The protocol developed in this study will facilitate the clinical applicability of all iPSC-LBs in the treatment of liver diseases. Nature Publishing Group UK 2020-10-21 /pmc/articles/PMC7578079/ /pubmed/33087763 http://dx.doi.org/10.1038/s41598-020-73908-1 Text en © The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Sekine, Keisuke
Ogawa, Shimpei
Tsuzuki, Syusaku
Kobayashi, Tatsuya
Ikeda, Kazuki
Nakanishi, Noriko
Takeuchi, Kenta
Kanai, Eriko
Otake, Yugo
Okamoto, Satoshi
Kobayashi, Tsuyoshi
Takebe, Takanori
Taniguchi, Hideki
Generation of human induced pluripotent stem cell-derived liver buds with chemically defined and animal origin-free media
title Generation of human induced pluripotent stem cell-derived liver buds with chemically defined and animal origin-free media
title_full Generation of human induced pluripotent stem cell-derived liver buds with chemically defined and animal origin-free media
title_fullStr Generation of human induced pluripotent stem cell-derived liver buds with chemically defined and animal origin-free media
title_full_unstemmed Generation of human induced pluripotent stem cell-derived liver buds with chemically defined and animal origin-free media
title_short Generation of human induced pluripotent stem cell-derived liver buds with chemically defined and animal origin-free media
title_sort generation of human induced pluripotent stem cell-derived liver buds with chemically defined and animal origin-free media
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7578079/
https://www.ncbi.nlm.nih.gov/pubmed/33087763
http://dx.doi.org/10.1038/s41598-020-73908-1
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