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Preserved in vivo reconstitution ability of PBSCs cryopreserved for a decade at −80 °C

Bone Marrow Transplantation volume 50, pages 1195–1200 (2015)Cite this article

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Abstract

PBSC products for auto- and allografting can be cryopreserved in liquid nitrogen with controlled-rate freezing until their use. Alternatively, they can be stored at −80 °C in a mechanical chest freezer, but it remains to be clarified whether PBSCs can be stored for the long term. We evaluated viability and functions of PBSCs cryopreserved for more than 10 years with this simplified method. Although recovery rate and viability of CD34+ cells were significantly decreased, myeloid differentiation potential and in vivo reconstitution and self-renewal potential of CD34+ cells in a xenogeneic engraftment assay were maintained for more than 10 years. These results indicate that PBSCs can be stored at −80 °C for years. Although accumulation of clinical engraftment data is required to confirm our results, this simplified cryopreservation will thus meet the increasing worldwide demand for PBSC transplantation in a region with limited resources.

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References

  1. Gratwohl A, Baldomero H, Gratwohl M, Aljurf M, Bouzas LF, Horowitz M et al. Quantitative and qualitative differences in use and trends of hematopoietic stem cell transplantation: a Global Observational Study. Haematologica 2013; 98: 1282–1290.

    Article  PubMed  PubMed Central  Google Scholar 

  2. Leemhuis T, Padley D, Keever-Taylor C, Niederwieser D, Teshima T, Lanza F et al. Essential requirements for setting up a stem cell processing laboratory. Bone Marrow Transplant 2014; 49: 1098–1105.

    Article  PubMed  Google Scholar 

  3. Stiff PJ, Murgo AJ, Zaroulis CG, DeRisi MF, Clarkson BD . Unfractionated human marrow cell cryopreservation using dimethylsulfoxide and hydroxyethyl starch. Cryobiology 1983; 20: 17–24.

    Article  CAS  PubMed  Google Scholar 

  4. Makino S, Harada M, Akashi K, Taniguchi S, Shibuya T, Inaba S et al. A simplified method for cryopreservation of peripheral blood stem cells at −80 degrees C without rate-controlled freezing. Bone Marrow Transplant 1991; 8: 239–244.

    CAS  PubMed  Google Scholar 

  5. Katayama Y, Yano T, Bessho A, Deguchi S, Sunami K, Mahmut N et al. The effects of a simplified method for cryopreservation and thawing procedures on peripheral blood stem cells. Bone Marrow Transplant 1997; 19: 283–287.

    Article  CAS  PubMed  Google Scholar 

  6. Ratajczak J, Marlicz W, Rozmyslowicz T, Machalinski B, Ratajczak MZ . Comparison of the different strategies for cryopreserving and storage of the bone marrow CD34+ cells. Possibility of unprogrammed rate freezing and storage at −80 degrees C mechanical freezer. Ann Transplant 1996; 1: 35–38.

    CAS  PubMed  Google Scholar 

  7. McCullough J, Haley R, Clay M, Hubel A, Lindgren B, Moroff G . Long-term storage of peripheral blood stem cells frozen and stored with a conventional liquid nitrogen technique compared with cells frozen and stored in a mechanical freezer. Transfusion 2010; 50: 808–819.

    Article  PubMed  Google Scholar 

  8. Watts MJ, Sullivan AM, Ings SJ, Barlow M, Devereux S, Goldstone AH et al. Storage of PBSC at −80 degrees C. Bone Marrow Transplant 1998; 21: 111–112.

    Article  CAS  PubMed  Google Scholar 

  9. Teshima T, Harada M, Takamatsu Y, Makino K, Taniguchi S, Inaba S et al. Cytotoxic drug and cytotoxic drug/G-CSF mobilization of peripheral blood stem cells and their use for autografting. Bone Marrow Transplant 1992; 10: 215–220.

    CAS  PubMed  Google Scholar 

  10. Shima T, Forraz N, Sato N, Yamauchi T, Iwasaki H, Takenaka K et al. A novel filtration method for cord blood processing using a polyester fabric filter. Int J Lab Hematol 2013; 35: 436–446.

    Article  CAS  PubMed  Google Scholar 

  11. Collison LW, Vignali DA . In vitro Treg suppression assays. Methods Mol Biol 2011; 707: 21–37.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Niiro H, Jabbarzadeh-Tabrizi S, Kikushige Y, Shima T, Noda K, Ota S et al. CIN85 is required for Cbl-mediated regulation of antigen receptor signaling in human B cells. Blood 2012; 119: 2263–2273.

    Article  CAS  PubMed  Google Scholar 

  13. Wang XM, Terasaki PI, Rankin GW Jr., Chia D, Zhong HP, Hardy S . A new microcellular cytotoxicity test based on calcein AM release. Hum Immunol 1993; 37: 264–270.

    Article  CAS  PubMed  Google Scholar 

  14. Fernyhough LJ, Buchan VA, McArthur LT, Hock BD . Relative recovery of haematopoietic stem cell products after cryogenic storage of up to 19 years. Bone Marrow Transplant 2013; 48: 32–35.

    Article  CAS  PubMed  Google Scholar 

  15. Liseth K, Ersvaer E, Abrahamsen JF, Nesthus I, Ryningen A, Bruserud O . Long-term cryopreservation of autologous stem cell grafts: a clinical and experimental study of hematopoietic and immunocompetent cells. Transfusion 2009; 49: 1709–1719.

    Article  CAS  PubMed  Google Scholar 

  16. Abbruzzese L, Agostini F, Durante C, Toffola RT, Rupolo M, Rossi FM et al. Long term cryopreservation in 5% DMSO maintains unchanged CD34(+) cells viability and allows satisfactory hematological engraftment after peripheral blood stem cell transplantation. Vox Sang 2013; 105: 77–80.

    Article  CAS  PubMed  Google Scholar 

  17. Costa R, Khattab M, Gilmore GL, Sahovic EA, Miller SM, Rossetti JM et al. Viability and potency of hematopoietic progenitor cells after prolonged cryopreservation at -80 degrees C. Acta Haematol 2011; 126: 99–102.

    Article  PubMed  Google Scholar 

  18. Detry G, Calvet L, Straetmans N, Cabrespine A, Ravoet C, Bay JO et al. Impact of uncontrolled freezing and long-term storage of peripheral blood stem cells at −80 degrees C on haematopoietic recovery after autologous transplantation. Report from two centres. Bone Marrow Transplant 2014; 49: 780–785.

    Article  CAS  PubMed  Google Scholar 

  19. Calvet L, Cabrespine A, Boiret-Dupre N, Merlin E, Paillard C, Berger M et al. Hematologic, immunologic reconstitution, and outcome of 342 autologous peripheral blood stem cell transplantations after cryopreservation in a −80 degrees C mechanical freezer and preserved less than 6 months. Transfusion 2013; 53: 570–578.

    Article  CAS  PubMed  Google Scholar 

  20. Iannalfi A, Bambi F, Tintori V, Lacitignola L, Bernini G, Mariani MP et al. Peripheral blood progenitor uncontrolled-rate freezing: a single pediatric center experience. Transfusion 2007; 47: 2202–2206.

    Article  PubMed  Google Scholar 

  21. Galmes A, Gutierrez A, Sampol A, Canaro M, Morey M, Iglesias J et al. Long-term hematological reconstitution and clinical evaluation of autologous peripheral blood stem cell transplantation after cryopreservation of cells with 5% and 10% dimethylsulfoxide at −80 degrees C in a mechanical freezer. Haematologica 2007; 92: 986–989.

    Article  PubMed  Google Scholar 

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Acknowledgements

This study was partly supported by grants from and Japan Agency for Medical Research and Development (AMED, 15545148).

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Authors and Affiliations

  1. Department of Medicine and Biosystemic Science, Kyushu University Graduate School of Medicine, Fukuoka, Japan

    T Shima, T Yamauchi, T Miyamoto & K Akashi

  2. Center for Cellular and Molecular Medicine, Kyushu University Graduate School of Medical Science, Fukuoka, Japan

    H Iwasaki, K Takenaka & K Akashi

  3. Clinical Chemistry and Laboratory Medicine, Kyushu University Graduate School of Medical Science, Fukuoka, Japan

    M Kadowaki, M Kiyosuke & T Mochimaru

  4. Department of Hematology, Hokkaido University Graduate School of Medicine, Sapporo, Japan

    T Teshima

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  1. T Shima
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Correspondence to T Teshima.

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Shima, T., Iwasaki, H., Yamauchi, T. et al. Preserved in vivo reconstitution ability of PBSCs cryopreserved for a decade at −80 °C. Bone Marrow Transplant 50, 1195–1200 (2015). https://doi.org/10.1038/bmt.2015.147

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  • DOI: https://doi.org/10.1038/bmt.2015.147

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