Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

Graft Manipulation

Impact of uncontrolled freezing and long-term storage of peripheral blood stem cells at −80 °C on haematopoietic recovery after autologous transplantation. Report from two centres

Bone Marrow Transplantation volume 49, pages 780–785 (2014)Cite this article

Subjects

Abstract

Controlled-rate freezing and storage in vapour phase nitrogen are used by most transplantation teams for the cryopreservation and storage of peripheral blood haematopoietic stem cells (PBSC). In this study, we analysed 666 autologous PBSC transplants after uncontrolled freezing and storage of PBSC at −80 °C. Statistical analysis showed that neutrophil recovery was associated with both the infused CD34+ cell dose (P=0.01) and the post transplantation use of growth factors (P<0.001) and that platelet recovery was associated with the infused CD34+ cell dose (P<0.001) and with the diagnosis (P=0.02). We analysed three groups according to the duration of the cryopreservation period (less than 6 months, between 6 and 12 months or more than 1 year). Haematopoietic recovery was not found to be adversely affected by longer storage at −80 °C. The haematopoietic recoveries of 50 pairs of sequential transplantations from the same PBSC mobilization were analysed. Despite prolonged cryopreservation, there were no statistically significant differences in neutrophil (P=0.09) or platelet (P=0.22) recovery in the second compared with the first transplant. In conclusion, the long-term storage of PBSC at −80 °C after uncontrolled-rate freezing is an easy and comparatively inexpensive cryopreservation method that leads to successful haematopoietic recovery even after prolonged storage.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1

Similar content being viewed by others

References

  1. Tedder RS, FFuckerman MA, Goldstone AH, Hawkins AE, Fielding A, Briggs EM et al. Hepatitis B transmission from contaminated cryopreservation tank. Lancet 1995; 346: 137–140.

    Article  CAS  Google Scholar 

  2. Balint B, Ivanović Z, Petakov M, Taseski J, Jovcić G, Stojanović N et al. The cryopreservation protocol optimal for progenitor recovery is not optimal for preservation of marrow repopulating ability. Bone Marrow Transplant 1999; 23: 613–619.

    Article  CAS  Google Scholar 

  3. Fleming KK, Hubel A . Cryopreservation of hematopoietic and non-hematopoietic stem cells. Transfus Apher Sci 2006; 34: 309–315.

    Article  CAS  Google Scholar 

  4. Akkok CA, Liseth K, Melve GK, Ersvaer E, Hervig T, Bruserud O . Is there a scientific basis for a recommended standardization of collection and cryopreservation of peripheral blood stem cell grafts? Cytotherapy 2011; 13: 1013–1024.

    Article  Google Scholar 

  5. 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  Google Scholar 

  6. Stiff PJ, Koester AR, Weidner MK, Dvorak K, Fisher RI . Autologous bone marrow transplantation using unfractionated cells cryopreserved in dimethylsulfoxide and hydroxyethyl starch without controlled-rate freezing. Blood 1987; 70: 974–978.

    CAS  PubMed  Google Scholar 

  7. Clark J, Pati A, McCarthy D . Successful cryopreservation of human bone marrow does not require a controlled-rate freezer. Bone Marrow Transplant 1991; 7: 121–125.

    CAS  PubMed  Google Scholar 

  8. 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 

  9. Rosenfeld CS, Gremba C, Shadduck RK, Zeigler ZR, Nemunaitis J . Engraftment with peripheral blood stem cells using noncontrolled-rate cryopreservation: comparison with autologous bone marrow transplantation. Exp Hematol 1994; 22: 290–294.

    CAS  PubMed  Google Scholar 

  10. Feremans WW, Bastin G, Moine FL, Ravoet C, Delville JP, Pradier O et al. Simplification of the blood stem cell transplantation (BSCT) procedure: large volume apheresis and uncontrolled rate cryopreservation at −80 degrees C. Eur J Haematol 1996; 56: 278–282.

    Article  CAS  Google Scholar 

  11. Choi CW, Kim BS, Seo JH, Shin SW, Kim YH, Kim JS . Long-term engraftment stability of peripheral blood stem cells cryopreserved using the dump-freezing method in a −80 degrees C mechanical freezer with 10% dimethyl sulfoxide. Int J Hematol 2001; 73: 245–250.

    Article  CAS  Google Scholar 

  12. Kudo Y, Minegishi M, Itoh T, Miura J, Saito N, Takahashi H et al. Evaluation of hematological reconstitution potential of autologous peripheral blood progenitor cells cryopreserved by a simple controlled-rate freezing method. Exp Med 2005; 205: 37–43.

    Google Scholar 

  13. Galmes A, Besalduch J, Bargay J, Matamoros N, Morey M, Novo A et al. A simplified method for cryopreservation of hematopoietic stem cells with −80 degrees C mechanical freezer with dimethyl sulfoxide as the sole cryoprotectant. Leuk Lymphoma 1995; 17: 181–184.

    Article  CAS  Google Scholar 

  14. Galmes A, Besalduch J, Bargay J, Matamoros N, Durán MA, Morey M et al. Cryopreservation of hematopoietic progenitor cells with 5-percent dimethyl sulfoxide at −80 degrees C without rate-controlled freezing. Transfusion 1996; 36: 794–797.

    Article  CAS  Google Scholar 

  15. Galmes A, Besalduch J, Bargay J, Novo A, Morey M, Guerra JM et al. Long-term storage at −80 degrees C of hematopoietic progenitor cells with 5-percent dimethyl sulfoxide as the sole cryoprotectant. Transfusion 1999; 39: 70–73.

    Article  CAS  Google Scholar 

  16. Galmes A, Gutiérrez 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  Google Scholar 

  17. Halle P, Tournilhac O, Knopinska-Posluszny W, Kanold J, Gembara P, Boiret N et al. Uncontrolled-rate freezing and storage at −80 degrees C, with only 3.5-percent DMSO in cryoprotective solution for 109 autologous peripheral blood progenitor cell transplantations. Transfusion 2001; 41: 667–673.

    Article  CAS  Google Scholar 

  18. Calvet L, Cabrespine A, Boiret-Dupré 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 °C mechanical freezer and preserved less than 6 months. Transfusion 2013; 53: 570–578.

    Article  CAS  Google Scholar 

  19. Aird W, Labopin M, Gorin NC, Antin JH . Long-term cryopreservation of human stem cells. Bone Marrow Transplant 1992; 9: 487–490.

    CAS  PubMed  Google Scholar 

  20. Motta MR, Benini C, Bandini G, Gherlinzoni F, Miggiano MC, Calori E et al. Autologous bone marrow transplantation with marrow cryopreserved for ten years. Bone Marrow Transplant 1993; 12: 177.

    CAS  PubMed  Google Scholar 

  21. Spurr EE, Wiggins NE, Marsden KA, Lowenthal RM, Ragg SJ . Cryopreserved human haematopoietic stem cells retain engraftment potential after extended (5-14 years) cryostorage. Cryobiology 2002; 44: 210–217.

    Article  Google Scholar 

  22. 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: 111–122.

    Article  CAS  Google Scholar 

  23. 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  Google Scholar 

  24. Kwiatkowski F, Girard M, Hacene K, Berlie J . Sem: a suitable statistical software adaptated for research in oncology. Bull Cancer 2000; 87: 715–721.

    CAS  PubMed  Google Scholar 

  25. Ayello J, Semidei-Pomales M, Preti R, Hesdorffer C, Reiss RF . Effects of long-term storage at −90 degrees C of bone marrow and PBPC on cell recovery, viability, and clonogenic potential. J Hematother 1998; 7: 385–390.

    Article  CAS  Google Scholar 

  26. Muramaki M, Hara I, Miyake H, Yamada Y, Okada H, Kamidono S . Long-term cryopreservation of peripheral blood stem cells in patients with advanced germ cell tumors using the dump-freezing method at −80 degrees C. Oncol Rep 2003; 10: 1993–1998.

    PubMed  Google Scholar 

  27. Hernandez-Navarro F, Ojeda E, Arrieta R, Rios-Rull P, Garcia-Bustos J, Quevedo E et al. Hematopoietic cell transplantation using plasma and DMSO without HES, with non-programmed freezing by immersion in a methanol bath: results in 213 cases. Bone Marrow Transplant 1998; 21: 511–517.

    Article  CAS  Google Scholar 

  28. Perez-Oteyza J, Bornstein R, Corral M, Hermosa V, Alegre A, Torrabadella M et al. Controlled-rate versus uncontrolled-rate cryopreservation of peripheral blood progenitor cells: a prospective multicenter study. Group for Cryobiology and Biology of Bone Marrow Transplantation (CBTMO), Spain. Haematologica 1998; 83: 1001–1005.

    CAS  PubMed  Google Scholar 

  29. Weaver CH, Hazelton B, Birch R, Palmer P, Allen C, Schwartzberg L et al. An analysis of engraftment kinetics as a function of the CD34 content of peripheral blood progenitor cell collections in 692 patients after the administration of myeloablative chemotherapy. Blood 1995; 86: 3961–3969.

    CAS  Google Scholar 

  30. Montanari M, Capelli D, Poloni A, Massidda D, Brunori M, Spitaleri L et al. A. Long-term hematologic reconstitution after autologous peripheral blood progenitor cell transplantation: a comparison between controlled-rate freezing and uncontrolled-rate freezing at 80 degrees C. Transfusion 2003; 43: 42–49.

    Article  Google Scholar 

  31. Feugier P, Bensoussan D, Girard F, Alla F, Schuhmacher A, Latger-Cannard V et al. Hematologic recovery after autologous PBPC transplantation: importance of the number of postthaw CD34+ cells. Transfusion 2003; 43: 878–884.

    Article  Google Scholar 

Download references

Acknowledgements

No source of support (grants, equipment, drugs) was required to conduct this study.

Author information

Authors and Affiliations

  1. Laboratoire d’hématologie, Centre Hospitalier de Jolimont, Haine Saint Paul, Belgium

    G Detry, B Husson & L Boon-Falleur

  2. Service de Thérapie cellulaire et Hématologie Clinique, Université d’Auvergne, CHU Estaing, Clermont-Ferrand, France

    L Calvet, A Cabrespine, J O Bay & O Tournilhac

  3. Service d’hématologie, Centre Hospitalier de Jolimont, Haine Saint Paul, Belgium

    N Straetmans, C Ravoet, H Petre & A Delannoy

  4. Centre régional de cancérologie et thérapie cellulaire pédiatrique, Centre de Biothérapie d’Auvergne, CHU Estaing, Clermont-Ferrand, France

    C Paillard, E Merlin & P Halle

Authors
  1. G Detry
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. L Calvet
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. N Straetmans
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. A Cabrespine
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. C Ravoet
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. J O Bay
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. H Petre
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. C Paillard
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. B Husson
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. E Merlin
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. L Boon-Falleur
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. O Tournilhac
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. A Delannoy
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. P Halle
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to G Detry.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Detry, G., Calvet, L., Straetmans, N. et al. Impact of uncontrolled freezing and long-term storage of peripheral blood stem cells at −80 °C on haematopoietic recovery after autologous transplantation. Report from two centres. Bone Marrow Transplant 49, 780–785 (2014). https://doi.org/10.1038/bmt.2014.53

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/bmt.2014.53

This article is cited by

Search

Advanced search

Quick links