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Suspension culture in a rotating bioreactor for efficient generation of human intestinal organoids
Human intestinal organoids (HIOs) derived from human pluripotent stem cells (hPSCs) hold great promise for translational medical applications. A common method to obtain HIOs has been to harvest floating hindgut spheroids arising from hPSCs. As this technique is elegant but burdensome due to the comp...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9701612/ https://www.ncbi.nlm.nih.gov/pubmed/36452871 http://dx.doi.org/10.1016/j.crmeth.2022.100337 |
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author | Takahashi, Junichi Mizutani, Tomohiro Sugihara, Hady Yuki Nagata, Sayaka Kato, Shu Hiraguri, Yui Takeoka, Sayaka Tsuchiya, Mao Kuno, Reiko Kakinuma, Sei Watanabe, Mamoru Okamoto, Ryuichi |
author_facet | Takahashi, Junichi Mizutani, Tomohiro Sugihara, Hady Yuki Nagata, Sayaka Kato, Shu Hiraguri, Yui Takeoka, Sayaka Tsuchiya, Mao Kuno, Reiko Kakinuma, Sei Watanabe, Mamoru Okamoto, Ryuichi |
author_sort | Takahashi, Junichi |
collection | PubMed |
description | Human intestinal organoids (HIOs) derived from human pluripotent stem cells (hPSCs) hold great promise for translational medical applications. A common method to obtain HIOs has been to harvest floating hindgut spheroids arising from hPSCs. As this technique is elegant but burdensome due to the complex protocol and line-to-line variability, a more feasible method is desired. Here, we establish a robust differentiation method into suspension-cultured HIOs (s-HIOs) by seeding dissociated cells on a spheroid-forming plate. This protocol realizes the reliable generation of size-controllable spheroids. Under optimized conditions in a rotating bioreactor, the generated spheroids quickly grow and mature into large s-HIOs with supporting mesenchyme. Upon mesenteric transplantation, s-HIOs further mature and develop complex tissue architecture in vivo. This method demonstrates that intestinal tissue can be generated from iPSC-derived HIOs via suspension induction and bioreactor maturation, establishing a reliable culture platform with wide applications in regenerative medicine. |
format | Online Article Text |
id | pubmed-9701612 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-97016122022-11-29 Suspension culture in a rotating bioreactor for efficient generation of human intestinal organoids Takahashi, Junichi Mizutani, Tomohiro Sugihara, Hady Yuki Nagata, Sayaka Kato, Shu Hiraguri, Yui Takeoka, Sayaka Tsuchiya, Mao Kuno, Reiko Kakinuma, Sei Watanabe, Mamoru Okamoto, Ryuichi Cell Rep Methods Report Human intestinal organoids (HIOs) derived from human pluripotent stem cells (hPSCs) hold great promise for translational medical applications. A common method to obtain HIOs has been to harvest floating hindgut spheroids arising from hPSCs. As this technique is elegant but burdensome due to the complex protocol and line-to-line variability, a more feasible method is desired. Here, we establish a robust differentiation method into suspension-cultured HIOs (s-HIOs) by seeding dissociated cells on a spheroid-forming plate. This protocol realizes the reliable generation of size-controllable spheroids. Under optimized conditions in a rotating bioreactor, the generated spheroids quickly grow and mature into large s-HIOs with supporting mesenchyme. Upon mesenteric transplantation, s-HIOs further mature and develop complex tissue architecture in vivo. This method demonstrates that intestinal tissue can be generated from iPSC-derived HIOs via suspension induction and bioreactor maturation, establishing a reliable culture platform with wide applications in regenerative medicine. Elsevier 2022-11-15 /pmc/articles/PMC9701612/ /pubmed/36452871 http://dx.doi.org/10.1016/j.crmeth.2022.100337 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Report Takahashi, Junichi Mizutani, Tomohiro Sugihara, Hady Yuki Nagata, Sayaka Kato, Shu Hiraguri, Yui Takeoka, Sayaka Tsuchiya, Mao Kuno, Reiko Kakinuma, Sei Watanabe, Mamoru Okamoto, Ryuichi Suspension culture in a rotating bioreactor for efficient generation of human intestinal organoids |
title | Suspension culture in a rotating bioreactor for efficient generation of human intestinal organoids |
title_full | Suspension culture in a rotating bioreactor for efficient generation of human intestinal organoids |
title_fullStr | Suspension culture in a rotating bioreactor for efficient generation of human intestinal organoids |
title_full_unstemmed | Suspension culture in a rotating bioreactor for efficient generation of human intestinal organoids |
title_short | Suspension culture in a rotating bioreactor for efficient generation of human intestinal organoids |
title_sort | suspension culture in a rotating bioreactor for efficient generation of human intestinal organoids |
topic | Report |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9701612/ https://www.ncbi.nlm.nih.gov/pubmed/36452871 http://dx.doi.org/10.1016/j.crmeth.2022.100337 |
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