Scale up manufacturing approach for production of human induced pluripotent stem cell-derived islets using Vertical Wheel® bioreactors
Abstract Advanced protocols show potential for human stem cells (SC)-derived islets generation under planar (2D) alone or three-dimensional (3D) cultures, but show challenges in scalability, cell loss, and batch-to-batch consistency. This study explores Vertical Wheel (VW)® bioreactor suspension tec...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2025-05-01
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| Series: | npj Regenerative Medicine |
| Online Access: | https://doi.org/10.1038/s41536-025-00409-y |
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| author | Nidheesh Dadheech Mario Bermúdez de León Zofia Czarnecka Nerea Cuesta-Gomez Ila Tewari Jasra Rena Pawlick Braulio Marfil-Garza Sandhya Sapkota Kevin Verhoeff Haide Razavy Perveen Anwar Abhineet Singh Nilanjan Ray Doug O’ Gorman Glen Jickling James Lyon Patrick MacDonald A. M. James Shapiro |
| author_facet | Nidheesh Dadheech Mario Bermúdez de León Zofia Czarnecka Nerea Cuesta-Gomez Ila Tewari Jasra Rena Pawlick Braulio Marfil-Garza Sandhya Sapkota Kevin Verhoeff Haide Razavy Perveen Anwar Abhineet Singh Nilanjan Ray Doug O’ Gorman Glen Jickling James Lyon Patrick MacDonald A. M. James Shapiro |
| author_sort | Nidheesh Dadheech |
| collection | DOAJ |
| description | Abstract Advanced protocols show potential for human stem cells (SC)-derived islets generation under planar (2D) alone or three-dimensional (3D) cultures, but show challenges in scalability, cell loss, and batch-to-batch consistency. This study explores Vertical Wheel (VW)® bioreactor suspension technology to differentiate islets from human induced pluripotent stem cells, achieving uniform, transcriptionally mature, and functional SC-islets. A 5x increase in scale from 0.1 L to 0.5 L reactors resulted in a 12-fold (15,005–183,002) increase in islet equivalent count (IEQ) without compromising islet structure. SC-islets show enriched β-cell composition (~63% CPPT+NKX6.1+ISL1+), glucose responsive insulin release (3.9–6.1-fold increase), and reversed diabetes in STZ-treated mice. Single cell RNA sequencing and flowcytometry analysis confirmed transcriptional maturity and functional identity, similar to adult islets. Lastly, harvested SC-islet grafts demonstrate improved islet functionality and mature transcriptomic signatures. Overall, scale-up in VW® bioreactor technology enhances IEQ yield with minimal variability and reduced cell loss, offering a pathway for clinical-grade SC-islet production. |
| format | Article |
| id | doaj-art-dbc52a3c4da44c108ccb7b2632f05966 |
| institution | OA Journals |
| issn | 2057-3995 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | npj Regenerative Medicine |
| spelling | doaj-art-dbc52a3c4da44c108ccb7b2632f059662025-08-20T02:39:03ZengNature Portfolionpj Regenerative Medicine2057-39952025-05-0110112310.1038/s41536-025-00409-yScale up manufacturing approach for production of human induced pluripotent stem cell-derived islets using Vertical Wheel® bioreactorsNidheesh Dadheech0Mario Bermúdez de León1Zofia Czarnecka2Nerea Cuesta-Gomez3Ila Tewari Jasra4Rena Pawlick5Braulio Marfil-Garza6Sandhya Sapkota7Kevin Verhoeff8Haide Razavy9Perveen Anwar10Abhineet Singh11Nilanjan Ray12Doug O’ Gorman13Glen Jickling14James Lyon15Patrick MacDonald16A. M. James Shapiro17Alberta Diabetes Institute, University of AlbertaCentro de Investigación Biomédica del Noreste, Departamento de Biología Molecular, Instituto Mexicano del Seguro SocialAlberta Diabetes Institute, University of AlbertaAlberta Diabetes Institute, University of AlbertaAlberta Diabetes Institute, University of AlbertaAlberta Diabetes Institute, University of AlbertaAlberta Diabetes Institute, University of AlbertaAlberta Diabetes Institute, University of AlbertaAlberta Diabetes Institute, University of AlbertaAlberta Diabetes Institute, University of AlbertaAlberta Diabetes Institute, University of AlbertaDepartment of Computing Science, University of AlbertaDepartment of Computing Science, University of AlbertaClinical Islet Transplant Program, University of AlbertaDepartment of Medicine, University of AlbertaAlberta Diabetes Institute, University of AlbertaAlberta Diabetes Institute, University of AlbertaAlberta Diabetes Institute, University of AlbertaAbstract Advanced protocols show potential for human stem cells (SC)-derived islets generation under planar (2D) alone or three-dimensional (3D) cultures, but show challenges in scalability, cell loss, and batch-to-batch consistency. This study explores Vertical Wheel (VW)® bioreactor suspension technology to differentiate islets from human induced pluripotent stem cells, achieving uniform, transcriptionally mature, and functional SC-islets. A 5x increase in scale from 0.1 L to 0.5 L reactors resulted in a 12-fold (15,005–183,002) increase in islet equivalent count (IEQ) without compromising islet structure. SC-islets show enriched β-cell composition (~63% CPPT+NKX6.1+ISL1+), glucose responsive insulin release (3.9–6.1-fold increase), and reversed diabetes in STZ-treated mice. Single cell RNA sequencing and flowcytometry analysis confirmed transcriptional maturity and functional identity, similar to adult islets. Lastly, harvested SC-islet grafts demonstrate improved islet functionality and mature transcriptomic signatures. Overall, scale-up in VW® bioreactor technology enhances IEQ yield with minimal variability and reduced cell loss, offering a pathway for clinical-grade SC-islet production.https://doi.org/10.1038/s41536-025-00409-y |
| spellingShingle | Nidheesh Dadheech Mario Bermúdez de León Zofia Czarnecka Nerea Cuesta-Gomez Ila Tewari Jasra Rena Pawlick Braulio Marfil-Garza Sandhya Sapkota Kevin Verhoeff Haide Razavy Perveen Anwar Abhineet Singh Nilanjan Ray Doug O’ Gorman Glen Jickling James Lyon Patrick MacDonald A. M. James Shapiro Scale up manufacturing approach for production of human induced pluripotent stem cell-derived islets using Vertical Wheel® bioreactors npj Regenerative Medicine |
| title | Scale up manufacturing approach for production of human induced pluripotent stem cell-derived islets using Vertical Wheel® bioreactors |
| title_full | Scale up manufacturing approach for production of human induced pluripotent stem cell-derived islets using Vertical Wheel® bioreactors |
| title_fullStr | Scale up manufacturing approach for production of human induced pluripotent stem cell-derived islets using Vertical Wheel® bioreactors |
| title_full_unstemmed | Scale up manufacturing approach for production of human induced pluripotent stem cell-derived islets using Vertical Wheel® bioreactors |
| title_short | Scale up manufacturing approach for production of human induced pluripotent stem cell-derived islets using Vertical Wheel® bioreactors |
| title_sort | scale up manufacturing approach for production of human induced pluripotent stem cell derived islets using vertical wheel r bioreactors |
| url | https://doi.org/10.1038/s41536-025-00409-y |
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