Protocol to generate large human intestinal organoids using a rotating bioreactor
Summary: Current techniques for producing induced-pluripotent-stem-cell-derived mid/hindgut spheroids have faced major hurdles in consistency and reproducibility. Here, we present a protocol that uses mid/hindgut cells to generate homogeneous spheroids that subsequently mature into human intestinal...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-09-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166723003416 |
| _version_ | 1797796334111031296 |
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| author | Junichi Takahashi Hady Yuki Sugihara Shu Kato Sayaka Nagata Ryuichi Okamoto Tomohiro Mizutani |
| author_facet | Junichi Takahashi Hady Yuki Sugihara Shu Kato Sayaka Nagata Ryuichi Okamoto Tomohiro Mizutani |
| author_sort | Junichi Takahashi |
| collection | DOAJ |
| description | Summary: Current techniques for producing induced-pluripotent-stem-cell-derived mid/hindgut spheroids have faced major hurdles in consistency and reproducibility. Here, we present a protocol that uses mid/hindgut cells to generate homogeneous spheroids that subsequently mature into human intestinal organoids (HIOs). We describe steps for stepwise differentiation and spheroid formation using a 96-well plate. We then detail cell maturation in a suspended state and the implementation of a rotational bioreactor platform to maximize the culture efficiency of larger HIOs.For complete details on the use and execution of this protocol, please refer to Takahashi et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-03-13T03:31:30Z |
| format | Article |
| id | doaj.art-8fbfcef431e44a2e99a841fee9adb792 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-03-13T03:31:30Z |
| publishDate | 2023-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-8fbfcef431e44a2e99a841fee9adb7922023-06-24T05:19:07ZengElsevierSTAR Protocols2666-16672023-09-0143102374Protocol to generate large human intestinal organoids using a rotating bioreactorJunichi Takahashi0Hady Yuki Sugihara1Shu Kato2Sayaka Nagata3Ryuichi Okamoto4Tomohiro Mizutani5Department of Gastroenterology and Hepatology, Tokyo Medical and Dental University (TMDU), 1-5-45, Yushima, Bunkyo-ku, Tokyo 113-8510, Japan; Corresponding authorDepartment of Gastroenterology and Hepatology, Tokyo Medical and Dental University (TMDU), 1-5-45, Yushima, Bunkyo-ku, Tokyo 113-8510, JapanDepartment of Gastroenterology and Hepatology, Tokyo Medical and Dental University (TMDU), 1-5-45, Yushima, Bunkyo-ku, Tokyo 113-8510, JapanDepartment of Gastroenterology and Hepatology, Tokyo Medical and Dental University (TMDU), 1-5-45, Yushima, Bunkyo-ku, Tokyo 113-8510, JapanDepartment of Gastroenterology and Hepatology, Tokyo Medical and Dental University (TMDU), 1-5-45, Yushima, Bunkyo-ku, Tokyo 113-8510, JapanDepartment of Gastroenterology and Hepatology, Tokyo Medical and Dental University (TMDU), 1-5-45, Yushima, Bunkyo-ku, Tokyo 113-8510, Japan; Corresponding authorSummary: Current techniques for producing induced-pluripotent-stem-cell-derived mid/hindgut spheroids have faced major hurdles in consistency and reproducibility. Here, we present a protocol that uses mid/hindgut cells to generate homogeneous spheroids that subsequently mature into human intestinal organoids (HIOs). We describe steps for stepwise differentiation and spheroid formation using a 96-well plate. We then detail cell maturation in a suspended state and the implementation of a rotational bioreactor platform to maximize the culture efficiency of larger HIOs.For complete details on the use and execution of this protocol, please refer to Takahashi et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166723003416Cell BiologyCell cultureStem CellsCell DifferentiationOrganoidsTissue Engineering |
| spellingShingle | Junichi Takahashi Hady Yuki Sugihara Shu Kato Sayaka Nagata Ryuichi Okamoto Tomohiro Mizutani Protocol to generate large human intestinal organoids using a rotating bioreactor STAR Protocols Cell Biology Cell culture Stem Cells Cell Differentiation Organoids Tissue Engineering |
| title | Protocol to generate large human intestinal organoids using a rotating bioreactor |
| title_full | Protocol to generate large human intestinal organoids using a rotating bioreactor |
| title_fullStr | Protocol to generate large human intestinal organoids using a rotating bioreactor |
| title_full_unstemmed | Protocol to generate large human intestinal organoids using a rotating bioreactor |
| title_short | Protocol to generate large human intestinal organoids using a rotating bioreactor |
| title_sort | protocol to generate large human intestinal organoids using a rotating bioreactor |
| topic | Cell Biology Cell culture Stem Cells Cell Differentiation Organoids Tissue Engineering |
| url | http://www.sciencedirect.com/science/article/pii/S2666166723003416 |
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