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:

In Vitro Studies

High sensitivity of megakaryocytic progenitor cells contained in placental/umbilical cord blood to the stresses during cryopreservation

Bone Marrow Transplantation volume 34, pages 537–543 (2004)Cite this article

Summary:

In placental/umbilical cord blood (PCB) banking and PCB transplantation (PCBT), long-term cryopreservation of hematopoietic stem and progenitor cells is a unique requirement as compared to that for bone marrow transplantation and cytokine-mobilized peripheral blood transplantation. A long period of severe thrombocytopenia is a problem in many patients after PCBT. The object of this study was to define whether megakaryocytic progenitor cells (CFU-Meg), which produce platelets, are more sensitive to cryopreservation than the other myeloid progenitor cells in PCB. The leukocyte concentrates (LCs) obtained from clinical PCB banks were cryopreserved, and progenitor cell recoveries were determined by differential count of colony-forming cells (CFCs). The LCs were exposed to stresses which cells face during freezing, thawing, and washing out cryoprotectants. Most of the myeloid progenitor cells contained in the LCs showed good survival when cryopreserved at slow cooling rates, although cellular injury was observed at higher cooling rates and higher osmolalities. In contrast, the recovery rate of CFU-Meg was significantly lower than other progenitor cells, indicating a higher sensitivity to the various stresses they were exposed to during cryopreservation. Thrombocytopenia observed in patients receiving PCBT may be explained, at least in part, by the disappearance of CFU-Meg during cryopreservation.

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
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6

Similar content being viewed by others

References

  1. Gluckman E, Broxmeyer HE, Auerbach AD et al. Hematopoietic reconstitution in a patient with Fanconi anemia by means of umbilical cord blood from an HLA-identical sibling. N Engl J Med 1989; 321: 1174–1178.

    Article  CAS  PubMed  Google Scholar 

  2. Rubinstein P, Rosenfield RE, Adamson JW, Stevens CE . Stored placental blood for unrelated bone marrow reconstitution. Blood 1993; 81: 1679–1690.

    CAS  PubMed  Google Scholar 

  3. Wagner JE, Kernan NA, Steinbuch M et al. Allogeneic sibling umbilical cord blood transplantation in children with malignant and non-malignant disease. Lancet 1995; 346: 214–219.

    Article  CAS  PubMed  Google Scholar 

  4. Kurtzberg J, Laughlin M, Graham ML et al. Placental blood as a source of hematopoietic stem cells for transplantation into unrelated recipients. N Engl J Med 1996; 335: 157–166.

    Article  CAS  PubMed  Google Scholar 

  5. Wagner JE, Rosenthal J, Sweetman R et al. Successful transplantation of HLA-matched and HLA-mismatched umbilical cord blood from unrelated donors: analysis of engraftment and acute graft-versus-host disease. Blood 1996; 88: 795–802.

    CAS  PubMed  Google Scholar 

  6. Gluckman E, Rocha V, Boyer-Chammard A et al. Outcome of cord-blood transplantation from related and unrelated donors. N Engl J Med 1997; 337: 373–381.

    Article  CAS  PubMed  Google Scholar 

  7. Rubinstein P, Dobrila L, Rosenfield RE et al. Processing and cryopreservation of placental/umbilical cord blood for unrelated bone marrow reconstitution. Proc Nat Acad Sci USA 1995; 92: 10119–10122.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Rubinstein P, Carrier C, Scaradavou A et al. Outcomes among 562 recipients of placental blood transplants from unrelated donors. N Engl J Med 1998; 339: 1565–1577.

    Article  CAS  PubMed  Google Scholar 

  9. Wagner JE . Umbilical cord transplantation. Leukemia 1998; 12 (Suppl. 1): S30–S32.

    PubMed  Google Scholar 

  10. Sirchia G, Rebulla P . Placental/umbilical cord blood transplantation. Haematologica 1999; 84: 738–747–747.

    CAS  PubMed  Google Scholar 

  11. Anasetti C, Amos D, Beatty PG et al. Effect of HLA compatibility on engraftment of bone marrow transplants in patients with leukemia or lymphoma. N Engl J Med 1989; 320: 197–204.

    Article  CAS  PubMed  Google Scholar 

  12. Kernan NA, Bartsch G, Ash RC et al. Analysis of 462 transplantations from unrelated donors facilitated by the National Marrow Donor Program. N Engl J Med 1993; 328: 593–602.

    Article  CAS  PubMed  Google Scholar 

  13. Balduzzi A, Gooley T, Anasetti C et al. Unrelated donor marrow transplantation in children. Blood 1995; 86: 3247–3256.

    CAS  PubMed  Google Scholar 

  14. Takahashi TA, Shibata K, Kashiwakura I et al. The effect of a hematopoietic-promoting factor (HPF) extracted from porcine kidney on the proliferation of human hematopoietic progenitor cells. Res Commun Mol Path 1996; 91: 185–193.

    CAS  Google Scholar 

  15. Kashiwakura I, Takahashi TA . Basic fibroblast growth factor-stimulated ex vivo expansion of human hematopoietic progenitor cells from placental and umbilical cord blood. Br J Haematol 2003; 122: 1–10.

    Article  Google Scholar 

  16. Kashiwakura I, Kuwabara M, Inanami O et al. Radiation sensitivity of megakaryocyte colony-forming cells in human placental/umbilical cord blood. Radiat Res 2000; 153: 144–152.

    Article  CAS  PubMed  Google Scholar 

  17. Siena S, Bregni M, Bonsi L et al. Increase in peripheral blood megakaryocyte progenitors following cancer therapy with high-dose cyclophosphamide and hematopoietic growth factors. Exp Hematol 1993; 21: 1583–1590.

    CAS  PubMed  Google Scholar 

  18. Hagiwara T, Kodama I, Horie K et al. Proliferative properties of human umbilical cord blood megakaryocyte progenitor cells to human thrombopoietin. Exp Hematol 1998; 26: 228–235.

    CAS  PubMed  Google Scholar 

  19. Handelsman H . Autologous peripheral stem-cell transplantation. Health Technol Assess (Rockv) 1995; 5: 1–16.

    Google Scholar 

  20. Woods EJ, Liu J, Derrow CW et al. Osmometric and permiability characteristics of human placental/umbilical cord blood CD34+ cells and their application to cryopreservation. J Hematother Stem Cell Res 2000; 9: 161–173.

    Article  CAS  PubMed  Google Scholar 

  21. Hunt CJ, Armitage SE, Pegg DE . Cryopreservation of umbilical cord blood: 2. Tolerance of CD34+ cells to multimolar dimethyl sulphoxide and the effect of cooling rate on recovery after freezing and thawing. Cryobiology 2003; 46: 76–87.

    Article  CAS  PubMed  Google Scholar 

  22. Crowe JH, Tablin F, Tsvetkova N et al. Are lipid phase transitions responsible for chilling damage in human platelets? Cryobiology 1999; 38: 180–191.

    Article  CAS  PubMed  Google Scholar 

  23. Drygalski A, Xu G, Constantinescu D et al. The frequency and proliferative potential of megakaryocytic colony-forming cells (Meg-CFC) in cord blood, cytokine-mobilized peripheral blood and bone marrow, and their correlation with total CFC numbers: implications for the quantitation of Meg-CFC to predict platelet engraftment following cord blood transplantation. Bone Marrow Transplant 2000; 25: 1029–1034.

    Article  CAS  PubMed  Google Scholar 

  24. Conrad PD, Emerson SG . Ex vivo expansion of hematopoietic cells from umbilical cord blood for clinical transplantation. J Leukocyte Biol 1998; 64: 147–155.

    Article  CAS  PubMed  Google Scholar 

  25. Gluckman E . Current status of umbilical cord blood hematopoietic stem cell transplantation. Exp Hematol 2000; 28: 1197–1205.

    Article  CAS  PubMed  Google Scholar 

  26. Demirer T, Ilhan O, Arat M et al. CD41+ and CD42+ hematopoietic progenitor cells may predict platelet engraftment after allogeneic peripheral blood stem cell transplantation. Clin Apheresis 2001; 16: 67–73.

    Article  CAS  Google Scholar 

  27. Begemann PG, Hassan HT, Kroger N et al. Correlation of time to platelet engraftment with amount of transplanted CD34+CD41+ cells after allogeneic bone marrow transplantation. J Hematother Stem Cell Res 2002; 11: 321–326.

    Article  CAS  PubMed  Google Scholar 

  28. Zauli G, Valvassori L, Capitani S . Presence and characteristics of circulating megakaryocyte progenitor cells in human fetal blood. Blood 1993; 81: 385–390.

    CAS  PubMed  Google Scholar 

  29. Bruno E, Murray LJ, DiGiusto R et al. Detection of a primitive megakaryocyte progenitor cell in human fetal bone marrow. Exp Hematol 1996; 24: 552–558.

    CAS  PubMed  Google Scholar 

  30. Miyazaki R, Ogata H, Iguchi T et al. Comparative analyses of megakaryocytes derived from cord blood and bone marrow. Br J Haematol 2000; 108: 602–609.

    Article  CAS  PubMed  Google Scholar 

  31. Pick M, Eldor A, Grisaru D et al. Ex vivo expansion of megakaryocyte progenitors from cryopreserved umbilical cord blood. A potential source of megakaryocytes for transplantation. Exp Hematol 2002; 30: 1079–1087.

    Article  PubMed  Google Scholar 

Download references

Acknowledgements

We thank Yasuo Tokushima, Makoto Enomoto, Mie Midorikawa, and Norihisa Sasayama for their special contribution to this study.

Author information

Authors and Affiliations

  1. Department of Cell and Development Biology, Vanderbilt University, Nashville, USA

    Y Xu

  2. Department of Radiological Technology, Hirosaki University School of Health Sciences, 66-1 Hon-cho, Hirosaki-shi, Aomori-ken, Japan

    I Kashiwakura

  3. Cell Processing Department, The Institute of Medical Science, The University of Tokyo, Shirokanedai 4-6-1, Minato-ku, Tokyo, Japan

    T A Takahashi

Authors
  1. Y Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. I Kashiwakura
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. T A Takahashi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to T A Takahashi.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Xu, Y., Kashiwakura, I. & Takahashi, T. High sensitivity of megakaryocytic progenitor cells contained in placental/umbilical cord blood to the stresses during cryopreservation. Bone Marrow Transplant 34, 537–543 (2004). https://doi.org/10.1038/sj.bmt.1704632

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.bmt.1704632

Keywords

Search

Advanced search

Quick links