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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...

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Autores principales: 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
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
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.
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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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