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Automated washing of long-term cryopreserved peripheral blood stem cells promotes cell viability and preserves CD34+ cell numbers

Bone Marrow Transplantation volume 53, pages 1225–1227 (2018)Cite this article

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Peripheral blood stem cell (PBSC) transplantation has become an established treatment option for a range of malignant and inherited diseases. PBSCs are usually cryopreserved in the presence of dimethyl sulfoxide (DMSO) and stored in liquid nitrogen. However, cryopreservation and thawing of PBSCs may affect cell viability, resulting in delayed engraftment and other risks related to low stem cell numbers [1]. DMSO is commonly used at a concentration of 10% during freezing of PBSCs. However, DMSO itself is toxic and lowering doses of DMSO may have a favorable effect on hematopoietic recovery after transplantation [2, 3]. Generally, PBSCs are thawed and infused without removal of DMSO. This has been associated with a wide range of adverse effects (AEs), ranging from minor to severe life-threatening events [4, 5]. Although not all toxic events can be contributed to DMSO, grafts containing lower concentrations of DMSO typically display a reduced incidence of AEs [5, 6]. To decrease AEs and improve graft quality after thawing, it has been suggested to remove DMSO prior to transplantation [4, 7] using different washing systems [8, 9]. Here, we aimed to remove DMSO from long-term cryopreserved PBSCs using an automated, fully closed system and assessed effects on CD34+ stem cells, viability, and colony-forming capacity.

PBSCs were collected from patients and healthy donors (n = 12), aged 46.8 ± 12.9 years, according to the standardized procedures. Ethical permission no. GO15/352-I8 was obtained from the internal Ethical Review Board of the Hacettepe University. PBSCs were collected using Fenwal Amicus (Fresenius Kabi, IL, USA) in a total volume of 60 mL, consisting of 30% cells and 70% plasma and Heparin (H3393, Sigma-Aldrich). Cells were frozen in 10% DMSO and stored for 3.9 ± 2.0 years in liquid nitrogen. Stem cell collection bags (OriGen Biomedical, Cryostore CS500CN, TX, USA) were rapidly thawed and placed on ice. To remove DMSO, the cell suspensions were washed with a fully automated, closed system (Biosafe, Sepax2, Switzerland) using 500 mL Voluven™ (Pfizer, USA), while maintaining the volume at 60 mL.

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Acknowledgements

This study was supported by a grant from the Hacettepe University Scientific Research Program Coordination Unit, project number EOZ_2016_7263.

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Authors and Affiliations

  1. Department of Stem Cell Sciences, Center for Stem Cell Research and Development, Hacettepe University Graduate School of Health Sciences, Ankara, Turkey

    Fatima Aerts-Kaya, Parisa Sharafi & Duygu Uçkan-Çetinkaya

  2. Department of Medical Oncology, Hacettepe University Cancer Institute, Ankara, Turkey

    Gül Koca & Funda Çakmak Sayla

  3. Department of Medical Oncology, Medicana International Hospital, Ankara, Turkey

    Evren Özdemir

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  1. Fatima Aerts-Kaya
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Correspondence to Fatima Aerts-Kaya.

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Aerts-Kaya, F., Koca, G., Sharafi, P. et al. Automated washing of long-term cryopreserved peripheral blood stem cells promotes cell viability and preserves CD34+ cell numbers. Bone Marrow Transplant 53, 1225–1227 (2018). https://doi.org/10.1038/s41409-018-0192-7

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  • DOI: https://doi.org/10.1038/s41409-018-0192-7

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