DMSO REMOVAL IN CRYOPRESERVED HEMATOPOIETIC PROGENITOR CELLS: PROCESS VALIDATION AND IMPACT ON CELL VIABILITY AND RECOVERY
- 1Grupo Pulsa São Paulo, São Paulo, SP, Brazil
Abstract
Introduction: Dimethyl sulfoxide (DMSO) is widely used as a cryoprotectant for the preservation of hematopoietic progenitor cells (HPCs). However, infusion of DMSO-containing products may be associated with adverse reactions, including cardiovascular, gastrointestinal, and neurological toxicities, particularly in pediatric patients and individuals with comorbidities. To minimize these effects, DMSO removal before infusion has been adopted in several cell processing laboratories. Nevertheless, washing and thawing procedures may negatively affect cell viability and recovery, potentially compromising product quality. Therefore, validation of DMSO removal protocols is essential to ensure safety, reproducibility, and therapeutic efficacy in hematopoietic cell transplantation. This study aimed to validate a standardized DMSO removal process in cryopreserved HPC products and evaluate its impact on cell viability and recovery. Methods: This validation study was conducted at our Cell Processing Center using cryopreserved HPC products discarded from clinical use after medical authorization. Samples originated from peripheral blood and were cryopreserved according to institutional protocols. Products were submitted to a standardized thawing and washing procedure using a solution containing 5% albumin and 6% hydroxyethyl starch (HES). Centrifugation was performed at 1400 rpm, 10°C, for 20 minutes. Total nucleated cell (TNC) count, cell viability, and percentage of cell recovery were evaluated before and after processing. Acceptance criteria were defined as viability ≥ 80% and cell recovery ≥ 80% after the procedure. Results: A total of 15 cryopreserved HPC samples submitted to the DMSO reduction process were analyzed. Mean pre-procedure cell viability was 98.3% (94.2–100%), while mean post-procedure viability was 97.6% (92.1–99.5%), demonstrating preservation of cellular integrity after processing, with a mean reduction below 1%. Mean cell recovery was 86.2% (SD ±9.7), with a median of 87.0% and values ranging from 68.8% to 100%. Eleven samples (73.3%) achieved cell recovery ≥ 80%, meeting the predefined validation criteria. Four samples showed recovery below 80%, suggesting cell loss possibly related to aggregate formation during washing and centrifugation. Most samples demonstrated satisfactory recovery and maintained high viability after processing. Conclusion: The DMSO removal process demonstrated adequate performance in preserving cell viability and maintaining satisfactory cellular recovery in most analyzed samples. The protocol proved reproducible and safe for processing cryopreserved HPC products, supporting its applicability as a strategy to reduce DMSO-related toxicity before infusion. These findings reinforce the importance of strict control of thawing and washing procedures to minimize cellular loss and ensure HPC product quality for hematopoietic transplantation.
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BibTeX
@article{calheiros2026dmso,
title = {{DMSO REMOVAL IN CRYOPRESERVED HEMATOPOIETIC PROGENITOR CELLS: PROCESS VALIDATION AND IMPACT ON CELL VIABILITY AND RECOVERY}},
author = {Waldely Veneziani CALHEIROS and Miria Lemos DA SILVA and Nayanna Faria DE CARVALHO and Erica Mioto Braz MERCHAN and Mirianceli Coelho MENDONÇA and Cesar De Almeida NETO},
journal = {Hematology, Transfusion and Cell Therapy},
year = {2026},
month = oct,
doi = {10.1016/j.htct.2026.106857},
url = {https://doi.org/10.1016/j.htct.2026.106857},
note = {CC BY},
}
RIS
TY - JOUR
TI - DMSO REMOVAL IN CRYOPRESERVED HEMATOPOIETIC PROGENITOR CELLS: PROCESS VALIDATION AND IMPACT ON CELL VIABILITY AND RECOVERY
AU - Waldely Veneziani CALHEIROS
AU - Miria Lemos DA SILVA
AU - Nayanna Faria DE CARVALHO
AU - Erica Mioto Braz MERCHAN
AU - Mirianceli Coelho MENDONÇA
AU - Cesar De Almeida NETO
PY - 2026
DA - 2026/10/23
JO - Hematology, Transfusion and Cell Therapy
DO - 10.1016/j.htct.2026.106857
UR - https://doi.org/10.1016/j.htct.2026.106857
AB - Introduction: Dimethyl sulfoxide (DMSO) is widely used as a cryoprotectant for the preservation of hematopoietic progenitor cells (HPCs). However, infusion of DMSO-containing products may be associated with adverse reactions, including cardiovascular, gastrointestinal, and neurological toxicities, particularly in pediatric patients and individuals with comorbidities. To minimize these effects, DMSO removal before infusion has been adopted in several cell processing laboratories. Nevertheless, washing and thawing procedures may negatively affect cell viability and recovery, potentially compromising product quality. Therefore, validation of DMSO removal protocols is essential to ensure safety, reproducibility, and therapeutic efficacy in hematopoietic cell transplantation. This study aimed to validate a standardized DMSO removal process in cryopreserved HPC products and evaluate its impact on cell viability and recovery. Methods: This validation study was conducted at our Cell Processing Center using cryopreserved HPC products discarded from clinical use after medical authorization. Samples originated from peripheral blood and were cryopreserved according to institutional protocols. Products were submitted to a standardized thawing and washing procedure using a solution containing 5% albumin and 6% hydroxyethyl starch (HES). Centrifugation was performed at 1400 rpm, 10°C, for 20 minutes. Total nucleated cell (TNC) count, cell viability, and percentage of cell recovery were evaluated before and after processing. Acceptance criteria were defined as viability ≥ 80% and cell recovery ≥ 80% after the procedure. Results: A total of 15 cryopreserved HPC samples submitted to the DMSO reduction process were analyzed. Mean pre-procedure cell viability was 98.3% (94.2–100%), while mean post-procedure viability was 97.6% (92.1–99.5%), demonstrating preservation of cellular integrity after processing, with a mean reduction below 1%. Mean cell recovery was 86.2% (SD ±9.7), with a median of 87.0% and values ranging from 68.8% to 100%. Eleven samples (73.3%) achieved cell recovery ≥ 80%, meeting the predefined validation criteria. Four samples showed recovery below 80%, suggesting cell loss possibly related to aggregate formation during washing and centrifugation. Most samples demonstrated satisfactory recovery and maintained high viability after processing. Conclusion: The DMSO removal process demonstrated adequate performance in preserving cell viability and maintaining satisfactory cellular recovery in most analyzed samples. The protocol proved reproducible and safe for processing cryopreserved HPC products, supporting its applicability as a strategy to reduce DMSO-related toxicity before infusion. These findings reinforce the importance of strict control of thawing and washing procedures to minimize cellular loss and ensure HPC product quality for hematopoietic transplantation.
SN - 2531-1379
SN - 2531-1387
ER -