Scale up manufacturing approach for production of human induced pluripotent stem cell-derived islets using Vertical Wheel® bioreactors
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<sup>+</sup>NKX6.1<sup>+</sup>ISL1<sup>+</sup>), 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.
Authors
- Nerea Cuesta‐Gomez (ORCID: https://orcid.org/0000-0002-1976-6298)
- Rena Pawlick (ORCID: https://orcid.org/0000-0002-0391-7901)
- Nidheesh Dadheech (ORCID: https://orcid.org/0000-0002-7155-0486)
- Patrick Edward MacDonald (ORCID: https://orcid.org/0000-0002-5439-6288)
- James Lyon (ORCID: https://orcid.org/0000-0001-7737-5183)
- Ila Tewari Jasra (ORCID: https://orcid.org/0000-0002-2842-8565)
- Kevin K. Verhoeff (ORCID: https://orcid.org/0000-0003-3110-5238)
- Mario Bermúdez de León (ORCID: https://orcid.org/0000-0002-0993-2969)
- Zofia Czarnecka (ORCID: https://orcid.org/0000-0002-8788-7585)
- A. M. James Shapiro (ORCID: https://orcid.org/0000-0002-6215-0990)
- Braulio A. Marfil‐Garza (ORCID: https://orcid.org/0000-0002-7224-2127)
- Haide Razavy (ORCID: https://orcid.org/0009-0000-5265-2965)
- Nilanjan Ray (ORCID: https://orcid.org/0000-0002-7588-5400)
- Glen C. Jickling (ORCID: https://orcid.org/0000-0003-3543-0820)
- Abhineet Singh (ORCID: https://orcid.org/0000-0002-5624-8377)
- Perveen Anwar
- Sandhya Sapkota
- Doug O’ Gorman
Institutions
- University of Alberta (CA)
- Mexican Social Security Institute (MX)
Publication Details
- Journal
- npj Regenerative Medicine
- Published
- 2025-05-29
- DOI
- https://doi.org/10.1038/s41536-025-00409-y
- Citations
- 6
- Primary Topic
- Pancreatic function and diabetes
- Type
- article
- Field-Weighted Citation Impact
- 12.04
Funders
- Diabetes Research Institute Foundation
- Juvenile Diabetes Research Foundation Canada
- Diabetes Canada
- Stem Cell Network
- Alberta Diabetes Foundation
- Canadian Institutes of Health Research
- Natural Sciences and Engineering Research Council of Canada