Scalable Bioreactor-based Suspension Approach to Generate Stem Cell-derived Islets From Healthy Donor-derived iPSCs.

Autor:	Verhoeff K; Alberta Diabetes Institute, University of Alberta, Edmonton, AB, Canada.; Department of Surgery, University of Alberta, Edmonton, AB, Canada., Cuesta-Gomez N; Alberta Diabetes Institute, University of Alberta, Edmonton, AB, Canada.; Department of Surgery, University of Alberta, Edmonton, AB, Canada.; Clinical Islet Transplant Program, University of Alberta, Edmonton, AB, Canada., Maghera J; Alberta Diabetes Institute, University of Alberta, Edmonton, AB, Canada.; Department of Pharmacology, University of Alberta, Edmonton, AB, Canada., Dadheech N; Alberta Diabetes Institute, University of Alberta, Edmonton, AB, Canada.; Department of Surgery, University of Alberta, Edmonton, AB, Canada.; Clinical Islet Transplant Program, University of Alberta, Edmonton, AB, Canada., Pawlick R; Alberta Diabetes Institute, University of Alberta, Edmonton, AB, Canada.; Department of Surgery, University of Alberta, Edmonton, AB, Canada.; Clinical Islet Transplant Program, University of Alberta, Edmonton, AB, Canada., Smith N; Alberta Diabetes Institute, University of Alberta, Edmonton, AB, Canada.; Clinical Islet Transplant Program, University of Alberta, Edmonton, AB, Canada.; Department of Pharmacology, University of Alberta, Edmonton, AB, Canada., O'Gorman D; Clinical Islet Transplant Program, University of Alberta, Edmonton, AB, Canada., Razavy H; Alberta Diabetes Institute, University of Alberta, Edmonton, AB, Canada.; Department of Surgery, University of Alberta, Edmonton, AB, Canada., Marfil-Garza B; Alberta Diabetes Institute, University of Alberta, Edmonton, AB, Canada.; National Institute of Medical Sciences and Nutrition Salvador Zubiran, Mexico City, Mexico.; CHRISTUS-LatAm Hub-Excellence and Innovation Center, Monterrey, Mexico., Young LG; ProtoKinetix Inc, Dalton, OH., Thiesen A; Department of Laboratory Medicine and Pathology, University of Alberta, Edmonton, AB, Canada., MacDonald PE; Alberta Diabetes Institute, University of Alberta, Edmonton, AB, Canada.; Department of Pharmacology, University of Alberta, Edmonton, AB, Canada., Shapiro AMJ; Alberta Diabetes Institute, University of Alberta, Edmonton, AB, Canada.; Department of Surgery, University of Alberta, Edmonton, AB, Canada.; Clinical Islet Transplant Program, University of Alberta, Edmonton, AB, Canada.
Jazyk:	angličtina
Zdroj:	Transplantation [Transplantation] 2024 Jul 18. Date of Electronic Publication: 2024 Jul 18.
DOI:	10.1097/TP.0000000000005108
Abstrakt:	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 × 104 cells/cm2 (total 8.3 × 106 cells), whereas complete suspension differentiation within 100 mL Vertical-Wheel bioreactors significantly increased cell yield to 1.1 × 106 cells/mL (total 105.0 × 106 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 enable future protocol comparison and evaluation of approaches for off-target cell elimination. Results suggest that bioreactor-based suspension differentiation protocols may facilitate scalability and clinical implementation of iPSC islet therapies. Competing Interests: A.M.J.S. serves as a consultant to ViaCyte Inc, Vertex Inc, Betalin Ltd, Hemostemix Inc, and Aspect Biosystems Ltd. A.M.J.S. is supported through a Canada Research Chair (tier 1) in Regenerative Medicine and Transplant Surgery and through grant support from the Juvenile Diabetes Research Foundation, Diabetes Canada, the Canadian Donation and Transplant Research Program, the Diabetes Research Institute Foundation of Canada, the Alberta Diabetes Foundation, and the Canadian Stem Cell Network. The other authors declare no conflicts of interest. (Copyright © 2024 Wolters Kluwer Health, Inc. All rights reserved.)
Databáze:	MEDLINE
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