Ice recrystallization inhibitors enable efficient cryopreservation of induced pluripotent stem cells: A functional and transcriptomic analysis

The successful use of human induced pluripotent stem cells (iPSCs) for research or clinical applications requires the development of robust, efficient, and reproducible cryopreservation protocols. After cryopreservation, the survival rate of iPSCs is suboptimal and cell line-dependent. We assessed t...

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Main Authors: Kathleen Mommaerts, Satoshi Okawa, Margaux Schmitt, Olga Kofanova, Tracey R. Turner, Robert N. Ben, Antonio Del Sol, William Mathieson, Jens C. Schwamborn, Jason P. Acker, Fay Betsou
Format: Article
Language:English
Published: Elsevier 2024-12-01
Series:Stem Cell Research
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Online Access:http://www.sciencedirect.com/science/article/pii/S1873506124002812
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author Kathleen Mommaerts
Satoshi Okawa
Margaux Schmitt
Olga Kofanova
Tracey R. Turner
Robert N. Ben
Antonio Del Sol
William Mathieson
Jens C. Schwamborn
Jason P. Acker
Fay Betsou
author_facet Kathleen Mommaerts
Satoshi Okawa
Margaux Schmitt
Olga Kofanova
Tracey R. Turner
Robert N. Ben
Antonio Del Sol
William Mathieson
Jens C. Schwamborn
Jason P. Acker
Fay Betsou
author_sort Kathleen Mommaerts
collection DOAJ
description The successful use of human induced pluripotent stem cells (iPSCs) for research or clinical applications requires the development of robust, efficient, and reproducible cryopreservation protocols. After cryopreservation, the survival rate of iPSCs is suboptimal and cell line-dependent. We assessed the use of ice recrystallization inhibitors (IRIs) for cryopreservation of human iPSCs. A toxicity screening study was performed to assess specific small-molecule carbohydrate-based IRIs and concentrations for further evaluation. Then, a cryopreservation study compared the cryoprotective efficiency of 15 mM IRIs in 5 % or 10 % DMSO-containing solutions and with CryoStor® CS10. Three iPSC lines were cryopreserved as single-cell suspensions in the cryopreservation solutions and post-thaw characteristics, including pluripotency and differential gene expression were assessed. We demonstrate the fitness-for-purpose of 15 mM IRI in 5 % DMSO as an efficient cryoprotective solution for iPSCs in terms of post-thaw recovery, viability, pluripotency, and transcriptomic changes. This mRNA sequencing dataset has the potential to be used for molecular mechanism analysis relating to cryopreservation. Use of IRIs can reduce DMSO concentrations and its associated toxicities, thereby improving the utility, effectiveness, and efficiency of cryopreservation.
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spelling doaj-art-f9fa9b08064a4d7591c9bfe26303d5932025-08-20T01:59:35ZengElsevierStem Cell Research1873-50612024-12-018110358310.1016/j.scr.2024.103583Ice recrystallization inhibitors enable efficient cryopreservation of induced pluripotent stem cells: A functional and transcriptomic analysisKathleen Mommaerts0Satoshi Okawa1Margaux Schmitt2Olga Kofanova3Tracey R. Turner4Robert N. Ben5Antonio Del Sol6William Mathieson7Jens C. Schwamborn8Jason P. Acker9Fay Betsou10Integrated Biobank of Luxembourg, Luxembourg Institute of Health, 1 rue Louis Rech, L-3555 Dudelange, Luxembourg; Luxembourg Centre for Systems Biomedicine (LCSB), University of Luxembourg, 2 avenue de Université, L-4365 Esch-sur-Alzette, Luxembourg; Corresponding author at: Integrated Biobank of Luxembourg (IBBL) - Luxembourg Institute of Health (LIH), 1, rue Louis Rech, L-3555 Dudelange, Luxembourg.Luxembourg Centre for Systems Biomedicine (LCSB), University of Luxembourg, 2 avenue de Université, L-4365 Esch-sur-Alzette, LuxembourgIntegrated Biobank of Luxembourg, Luxembourg Institute of Health, 1 rue Louis Rech, L-3555 Dudelange, LuxembourgIntegrated Biobank of Luxembourg, Luxembourg Institute of Health, 1 rue Louis Rech, L-3555 Dudelange, LuxembourgPanTHERA CryoSolutions Inc., Edmonton, Alberta, CanadaPanTHERA CryoSolutions Inc., Edmonton, Alberta, Canada; Department of Chemistry, University of Ottawa, Ottawa, Ontario, CanadaLuxembourg Centre for Systems Biomedicine (LCSB), University of Luxembourg, 2 avenue de Université, L-4365 Esch-sur-Alzette, Luxembourg; CIC bioGUNE, Bizkaia Technology Park, 48160 Derio, Spain; IKERBASQUE, Basque Foundation for Science, Bilbao 48013, SpainIntegrated Biobank of Luxembourg, Luxembourg Institute of Health, 1 rue Louis Rech, L-3555 Dudelange, LuxembourgLuxembourg Centre for Systems Biomedicine (LCSB), University of Luxembourg, 2 avenue de Université, L-4365 Esch-sur-Alzette, LuxembourgPanTHERA CryoSolutions Inc., Edmonton, Alberta, Canada; Laboratory Medicine and Pathology, University of Alberta, Edmonton, Alberta, CanadaIntegrated Biobank of Luxembourg, Luxembourg Institute of Health, 1 rue Louis Rech, L-3555 Dudelange, LuxembourgThe successful use of human induced pluripotent stem cells (iPSCs) for research or clinical applications requires the development of robust, efficient, and reproducible cryopreservation protocols. After cryopreservation, the survival rate of iPSCs is suboptimal and cell line-dependent. We assessed the use of ice recrystallization inhibitors (IRIs) for cryopreservation of human iPSCs. A toxicity screening study was performed to assess specific small-molecule carbohydrate-based IRIs and concentrations for further evaluation. Then, a cryopreservation study compared the cryoprotective efficiency of 15 mM IRIs in 5 % or 10 % DMSO-containing solutions and with CryoStor® CS10. Three iPSC lines were cryopreserved as single-cell suspensions in the cryopreservation solutions and post-thaw characteristics, including pluripotency and differential gene expression were assessed. We demonstrate the fitness-for-purpose of 15 mM IRI in 5 % DMSO as an efficient cryoprotective solution for iPSCs in terms of post-thaw recovery, viability, pluripotency, and transcriptomic changes. This mRNA sequencing dataset has the potential to be used for molecular mechanism analysis relating to cryopreservation. Use of IRIs can reduce DMSO concentrations and its associated toxicities, thereby improving the utility, effectiveness, and efficiency of cryopreservation.http://www.sciencedirect.com/science/article/pii/S1873506124002812Ice recrystallization inhibitorsInduced pluripotent stem cellsCryopreservationTranscriptomeDMSOPluripotency
spellingShingle Kathleen Mommaerts
Satoshi Okawa
Margaux Schmitt
Olga Kofanova
Tracey R. Turner
Robert N. Ben
Antonio Del Sol
William Mathieson
Jens C. Schwamborn
Jason P. Acker
Fay Betsou
Ice recrystallization inhibitors enable efficient cryopreservation of induced pluripotent stem cells: A functional and transcriptomic analysis
Stem Cell Research
Ice recrystallization inhibitors
Induced pluripotent stem cells
Cryopreservation
Transcriptome
DMSO
Pluripotency
title Ice recrystallization inhibitors enable efficient cryopreservation of induced pluripotent stem cells: A functional and transcriptomic analysis
title_full Ice recrystallization inhibitors enable efficient cryopreservation of induced pluripotent stem cells: A functional and transcriptomic analysis
title_fullStr Ice recrystallization inhibitors enable efficient cryopreservation of induced pluripotent stem cells: A functional and transcriptomic analysis
title_full_unstemmed Ice recrystallization inhibitors enable efficient cryopreservation of induced pluripotent stem cells: A functional and transcriptomic analysis
title_short Ice recrystallization inhibitors enable efficient cryopreservation of induced pluripotent stem cells: A functional and transcriptomic analysis
title_sort ice recrystallization inhibitors enable efficient cryopreservation of induced pluripotent stem cells a functional and transcriptomic analysis
topic Ice recrystallization inhibitors
Induced pluripotent stem cells
Cryopreservation
Transcriptome
DMSO
Pluripotency
url http://www.sciencedirect.com/science/article/pii/S1873506124002812
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