Scalable bioreactor-based suspension approach to generate stem cell-derived islets from healthy donor-derived iPSCs
Abstract
Background Induced pluripotent stem cells (iPSCs) offer the potential to generate autologous iPSC-derived islets (iPSC islets), however, remain limited by scalability and product safety. Methods Herein, we report stagewise characterization of cells generated following a bioreactor-based differentiation protocol. Cell characteristics were assessed using flow cytometry, quantitative reverse transcription polymerase chain reaction, patch clamping, functional assessment, and in vivo functional and immunohistochemistry evaluation. Protocol yield and costs are assessed to determine scalability. Results Differentiation was capable of generating 90.4% PDX1 + /NKX6.1 + pancreatic progenitors and 100% C-peptide + /NKX6.1 + iPSC islet cells. However, 82.1%, 49.6%, and 0.9% of the cells expressed SOX9 (duct), SLC18A1 (enterochromaffin cells), and CDX2 (gut cells), respectively. Explanted grafts contained mature monohormonal islet-like cells, however, CK19 + ductal tissues persist. Using this protocol, semi-planar differentiation using 150 mm plates achieved 5.72 × 10 4 cells/cm 2 (total 8.3 × 10 6 cells), whereas complete suspension differentiation within 100 mL Vertical-Wheel bioreactors significantly increased cell yield to 1.1 × 10 6 cells/mL (total 105.0 × 10 6 cells), reducing costs by 88.8%. Conclusions This study offers a scalable suspension-based approach for iPSC islet differentiation within Vertical-Wheel bioreactors with thorough characterization of the ensuing product to enab…
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Approximately 1 billion pancreatic progenitor cells are estimated to be required per person, although this number could be less if more mature cells are used.
“Approximately 1 billion pancreatic progenitor cells are estimated to be required per person, although this number could be less if more mature cells are used.”
From The future of type 1 diabetes therapy (2025) · cites this paper as [78]
Treating many people with type 1 diabetes will require substantial scale-up of cell manufacturing and other changes, such as potentially shortening the duration of in-vitro steps to reduce cost and increase efficiency.
“Treating many people with type 1 diabetes will require substantial scaleup of cell manufacturing and other changes, such as potentially shortening the duration of invitro steps to reduce cost and increase efficiency.78,79”
From The future of type 1 diabetes therapy (2025) · cites this paper as [78]
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- Authors
- Verhoeff, K., Cuesta-Gomez, N., Maghera, J., Dadheech, N., Pawlick, R., Smith, N., O’Gorman, D., Razavy, H., Marfil-Garza, B., Young, L. G., Thiesen, A., MacDonald, P. E., & Shapiro, A. M. J.
- Affiliations
- University of Alberta (CA)
- Alberta Diabetes Institute, University of Alberta, Edmonton, AB, Canada.
- ProtoKinetix Inc, Dalton, OH.
- Corresponding author
- Shapiro, A. M. James
- ORCID
- Venue
- Transplantation
- Published
- 2024-07-18
- Volume
- 109
- Issue
- 1
- Pages
- e22-e35
- ISSN
- 0041-1337 (ISSN-L)
- 1534-6080
- OpenAlex
- W4400797315
- Cited by
- 16
- References
- 0
- Citations by year
- 2026: 9 · 2025: 6 · 2024: 1
- FWCI
- 6.76
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Transplantation (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Breakthrough T1D Canada (5-SRA-2021-1148-S-B, 2-PAR-2023-1312-A-N)
- Topics
- Health Sciences › Medicine › Surgery › Pancreatic function and diabetes
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Diabetes and associated disorders
- Keywords
- induced pluripotent stem cells, vertical-wheel bioreactor, pancreatic progenitors, NKX6.1, PDX1, C-peptide, islet-like cells, islet transplantation, autologous cell therapy, cost reduction
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