Cryopreservation of mesenchymal stem/stromal cells using a DMSO-free solution is comparable to DMSO-containing cryoprotectants: results of an international multicenter PACT/BEST collaborative study.

Autor: Mamo T; Department of Laboratory Medicine and Pathology, University of Minnesota, Minneapolis, Minnesota, USA. Electronic address: mamo0016@umn.edu., Cox CA; Moffitt Cancer Center, Tampa, Florida, USA., Demorest C; Masonic Cancer Center Biostatistics Core, University of Minnesota, Minneapolis, Minnesota, USA., Fontaine MJ; University of Maryland School of Medicine, Baltimore, Maryland, USA., Hubel A; Department of Mechanical Engineering, University of Minnesota, Minneapolis, Minnesota, USA; Evia Bio, Minneapolis, Minnesota, USA., Kelley L; Moffitt Cancer Center, Tampa, Florida, USA., Khan A; University of Miami, Coral Gables, Florida, USA., Marks DC; Research and Development, Australian Red Cross Lifeblood, Sydney, Australia., Pati S; University of California San Francisco, San Francisco, California, USA., Reems JA; University of Utah, Salt Lake City, Utah, USA., Spohn G; German Red Cross Blood Donor Service and Goethe University Hospital, Frankfurt am Main, Germany., Schäfer R; German Red Cross Blood Donor Service and Goethe University Hospital, Frankfurt am Main, Germany; Medical Center, Center for Chronic Immunodeficiency, University of Freiburg, Freiburg, Germany., Shi R; Center for Cellular Engineering, Clinical Center, National Institutes of Health, Bethesda, Maryland, USA., Shao L; Center for Cellular Engineering, Clinical Center, National Institutes of Health, Bethesda, Maryland, USA., Stroncek D; Center for Cellular Engineering, Clinical Center, National Institutes of Health, Bethesda, Maryland, USA., McKenna DH; Department of Laboratory Medicine and Pathology, University of Minnesota, Minneapolis, Minnesota, USA; Molecular and Cellular Therapeutics, University of Minnesota, Saint Paul, Minnesota, USA.
Jazyk: angličtina
Zdroj: Cytotherapy [Cytotherapy] 2024 Jul 06. Date of Electronic Publication: 2024 Jul 06.
DOI: 10.1016/j.jcyt.2024.07.001
Abstrakt: Background and Aim: An essential aspect of ensuring availability and stability of mesenchymal stem/stromal cells (MSCs) products for clinical use is that these cells are cryopreserved before individual infusion into patients. Currently, cryopreservation of MSCs involves use of a cryoprotectant solution containing dimethyl sulfoxide (DMSO). However, it is recognized that DMSO may be toxic for both the patient and the MSC product. In this Production Assistance for Cellular Therapies (PACT) and Biomedical Excellence for Safer Transfusion (BEST) Collaborative study, we compared a novel DMSO-free solution with DMSO containing cryoprotectant solutions for freezing MSCs.
Methods: A DMSO-free cryoprotectant solution containing sucrose, glycerol, and isoleucine (SGI) in a base of Plasmalyte A was prepared at the University of Minnesota. Cryoprotectant solutions containing 5-10% DMSO (in-house) were prepared at seven participating centers (five from USA, one each from Australia and Germany). The MSCs were isolated from bone marrow or adipose tissue and cultured ex vivo per local protocols at each center. The cells in suspension were frozen by aliquoting into vials/bags. For six out of the seven centers, the vials/bags were placed in a controlled rate freezer (one center placed them at -80°C freezer overnight) before transferring to liquid nitrogen. The cells were kept frozen for at least one week before thawing and testing. Pre- and post-thaw assessment included cell viability and recovery, immunophenotype as well as transcriptional and gene expression profiles. Linear regression, mixed effects models and two-sided t-tests were applied for statistical analysis.
Results: MSCs had an average viability of 94.3% (95% CI: 87.2-100%) before cryopreservation, decreasing by 4.5% (95% CI: 0.03-9.0%; P: 0.049) and 11.4% (95% CI: 6.9-15.8%; P< 0.001), for MSCs cryopreserved in the in-house and SGI solutions, respectively. The average recovery of viable MSCs cryopreserved in the SGI was 92.9% (95% CI: 85.7-100.0%), and it was lower by 5.6% (95% CI: 1.3-9.8%, P < 0.013) for the in-house solution. Additionally, MSCs cryopreserved in the two solutions had expected level of expressions for CD45, CD73, CD90, and CD105 with no significant difference in global gene expression profiles.
Conclusion: MSCs cryopreserved in a DMSO-free solution containing sucrose, glycerol, and isoleucine in a base of Plasmalyte A had slightly lower cell viability, better recovery, and comparable immunophenotype and global gene expression profiles compared to MSCs cryopreserved in DMSO containing solutions. The average viability of MSCs in the novel solution was above 80% and, thus, likely clinically acceptable. Future studies are suggested to test the post-thaw functions of MSCs cryopreserved in the novel DMSO-free solution.
Competing Interests: Declaration of competing interest AH is the Founder and Chief Scientific Officer of Evia Bio, a company that was established after the current study was completed and is commercializing the proprietary SGI solution used in this study.
(Copyright © 2024 International Society for Cell & Gene Therapy. Published by Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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