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Cell Characterization, Graft Evaluation, and Yield of Islet-Like Cells Differentiated From Patient-Derived iPSCs

77 Pages Posted: 19 May 2023 Publication Status: Review Complete

See all articles by Kevin Verhoeff

Kevin Verhoeff

University of Alberta - Alberta Diabetes Institute

Nerea Cuesta-Gomez

University of Alberta - Alberta Diabetes Institute; University of Alberta - Department of Surgery

Jasmine Maghera

University of Alberta - Alberta Diabetes Institute

Nidheesh Dadheech

University of Alberta - Alberta Diabetes Institute

Rena Pawlick

University of Alberta - Alberta Diabetes Institute

Nancy Smith

University of Alberta - Alberta Diabetes Institute

Doug O’Gorman

University of Alberta - Clinical Islet Transplant Program

Haide Razavy

University of Alberta - Alberta Diabetes Institute

Braulio A. Marfil-Garza

University of Alberta - Alberta Diabetes Institute

Lachlan G. Young

ProtoKinetix Inc.

Patrick E. MacDonald

University of Alberta - Alberta Diabetes Institute

James Shapiro

University of Alberta - Alberta Diabetes Institute; University of Alberta - Department of Surgery

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Abstract

Induced pluripotent stem cells (iPSCs) offer the potential to generate autologous iPSC-derived islets (iPSC islets). Differentiation protocol optimization with stage-wise characterization, off-target evaluation, and cell yield assessment are necessary to inform clinical implementation. Herein, we report stage-wise characterization of cells generated following an improved differentiation protocol 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. Importantly, planar differentiation achieved 8.3x106+ cells, whereas complete suspension differentiation within Vertical-Wheel® bioreactors significantly increased cell yield to 105.0x106 cells, reducing costs by 88.8%. This study offers improved stage-wise characterization of iPSC islet cells that will enable future protocol comparison and evaluation of approaches for off-target cell elimination. Proof-of-concept for complete suspension-based differentiation highlights an important advancement to facilitate clinical implementation.

Note:

Funding Information: The work within this study was supported by grants 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.

Declaration of Interests: AMJS serves as a consultant to ViaCyte Inc., Vertex Inc., Betalin Ltd., Hemostemix Inc. and Aspect Biosystems Ltd. AMJS 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. Braulio A. Marfil–Garza is supported by the Patronato del Instituto Nacional de Ciencias Medicas y Nutricion Salvador Zubiran (INCMNSZ), the Fundacion para la Salud y la Educacion Dr. Salvador Zubirán (FunSaEd), and the CHRISTUS Excellence and Innovation Center. All other authors have no conflicts of interest to disclose.

Ethical Approval Statement: Blood sample donors for this study provided written consent for use of tissue, cell reprogramming and differentiation, and result disclosure. This study and its methods have been approved by the Stem Cell Oversight Committee of Canada (SCOC), and the University of Alberta Institutional Health Research Ethics Board (PRO00084032).

Animal protocols were conducted in accordance with the Canadian Council on Animal Care Guidelines and Policies and have been approved by the Animal Care and Use Committee (Health Sciences) at the University of Alberta.

Keywords: induced pluripotent stem cells, islet cell transplant, cell therapy, islet differentiation, personalized medicine, characterization, yield, scalability

Suggested Citation

Verhoeff, Kevin and Cuesta-Gomez, Nerea and Maghera, Jasmine and Dadheech, Nidheesh and Pawlick, Rena and Smith, Nancy and O’Gorman, Doug and Razavy, Haide and Marfil-Garza, Braulio A. and Young, Lachlan G. and MacDonald, Patrick E. and Shapiro, James, Cell Characterization, Graft Evaluation, and Yield of Islet-Like Cells Differentiated From Patient-Derived iPSCs. Available at SSRN: https://ssrn.com/abstract=4450374 or http://dx.doi.org/10.2139/ssrn.4450374
This version of the paper has not been formally peer reviewed.

Kevin Verhoeff

University of Alberta - Alberta Diabetes Institute ( email )

Nerea Cuesta-Gomez

University of Alberta - Alberta Diabetes Institute ( email )

University of Alberta - Department of Surgery ( email )

Jasmine Maghera

University of Alberta - Alberta Diabetes Institute ( email )

Nidheesh Dadheech

University of Alberta - Alberta Diabetes Institute ( email )

Rena Pawlick

University of Alberta - Alberta Diabetes Institute ( email )

Nancy Smith

University of Alberta - Alberta Diabetes Institute ( email )

Doug O’Gorman

University of Alberta - Clinical Islet Transplant Program ( email )

Haide Razavy

University of Alberta - Alberta Diabetes Institute ( email )

Braulio A. Marfil-Garza

University of Alberta - Alberta Diabetes Institute ( email )

Lachlan G. Young

ProtoKinetix Inc. ( email )

Patrick E. MacDonald

University of Alberta - Alberta Diabetes Institute ( email )

James Shapiro (Contact Author)

University of Alberta - Alberta Diabetes Institute ( email )

University of Alberta - Department of Surgery ( email )

Canada

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