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.

  • Letter to the Editor
  • Published:

Transplantation of human peripheral blood CD34-positive cells in combination with ex vivo generated megakaryocytes results in fast platelet formation in NOD/SCID mice

Leukemia volume 22, pages 203–208 (2008)Cite this article

Many types of cancers are currently treated with high-dose chemotherapy, which affects all (rapidly) dividing cells, including hematopoietic progenitor cells (HPCs). Therefore, even after stem cell transplantation, patients often suffer from a period of pancytopenia. Anemia and leukopenia, in this respect, can be successfully treated with Epo and G-CSF, respectively, by inducing a faster recovery of hemoglobin and neutrophil levels. Several attempts have been undertaken to shorten the period of thrombocytopenia by co-transplantation of autologous megakaryocytes (MKs) expanded from CD34+ HPCs, but the effect of this treatment was variable and if present, minimal.1, 2, 3, 4 However, the MK expansion protocols used in these clinical trials vary considerably and optimization of the composition of the transplant may therefore increase the clinical benefit of co-transplantation of expanded MKs. We therefore investigated whether MKs generated from mobilized peripheral blood (MPB) CD34+ could induce fast platelet production when combined with unmanipulated CD34+ cells after transplantation in the nonobese diabetic/severe combined immunodeficiency (NOD/SCID) mouse. For this purpose, CD34+ cells were cultured for 7 days with 100 ng ml−1 thrombopoietin (Tpo) and 10 ng ml−1 interleukin-1β (IL-1β), a cytokine cocktail, previously shown to efficiently generate MK progenitors and mature MKs (up to 32N) in our lab.5

The NOD/SCID mouse model has long been accepted as a model for human hematopoiesis.6 Transplantation of human MPB CD34+ cells can result in high levels of engraftment and differentiation along multiple lineages in the bone marrow (BM) of NOD/SCID mice.7 In two previous studies,8, 9 in which human platelet formation after transplantation of MPB CD34+ cells in NOD/SCID mice was investigated, the number of cells transplanted was 0.6–1 × 106 per mouse. Transplantation of such numbers of CD34+ cells resulted in a variable and relatively low level (<10%) of human CD45+ cells in the BM for most mice. Since the number of transplanted MPB CD34+ cells has been shown to correlate with the observed chimerism in the mouse BM and blood,7 we decided to transplant at least 2.25 × 106 unmanipulated MPB CD34+ cells per mouse in the groups that received combined grafts, to facilitate accurate measurement of human platelet formation.

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

References

  1. Bertolini F, Battaglia M, Pedrazzoli P, Da Prada GA, Lanza A, Soligo D et al. Megakaryocytic progenitors can be generated ex vivo and safely administered to autologous peripheral blood progenitor cell transplant recipients. Blood 1997; 89: 2679–2688.

    CAS  Google Scholar 

  2. Decaudin D, Vantelon JM, Bourhis JH, Farace F, Bonnet ML, Guillier M et al. Ex vivo expansion of megakaryocyte precursor cells in autologous stem cell transplantation for relapsed malignant lymphoma. Bone Marrow Transplant 2004; 34: 1089–1093.

    Article  CAS  Google Scholar 

  3. Prince HM, Simmons PJ, Whitty G, Wall DP, Barber L, Toner GC et al. Improved haematopoietic recovery following transplantation with ex vivo-expanded mobilized blood cells. Br J Haematol 2004; 126: 536–545.

    Article  Google Scholar 

  4. Scheding S, Bergmannn M, Rathke G, Vogel W, Brugger W, Kanz L . Additional transplantation of ex vivo generated megakaryocytic cells after high-dose chemotherapy. Haematologica 2004; 89: 630–631.

    Google Scholar 

  5. van den Oudenrijn S, de Haas M, Calafat J, van der Schoot CE, von dem Borne AE . A combination of megakaryocyte growth and development factor and interleukin-1 is sufficient to culture large numbers of megakaryocytic progenitors and megakaryocytes for transfusion purposes. Br J Haematol 1999; 106: 553–563.

    Article  CAS  Google Scholar 

  6. Lapidot T, Fajerman Y, Kollet O . Immune-deficient SCID and NOD/SCID mice models as functional assays for studying normal and malignant human hematopoiesis. J Mol Med 1997; 75: 664–673.

    Article  CAS  Google Scholar 

  7. van der Loo JC, Hanenberg H, Cooper RJ, Luo FY, Lazaridis EN, Williams DA . Nonobese diabetic/severe combined immunodeficiency (NOD/SCID) mouse as a model system to study the engraftment and mobilization of human peripheral blood stem cells. Blood 1998; 92: 2556–2570.

    CAS  Google Scholar 

  8. Angelopoulou MK, Rinder H, Wang C, Burtness B, Cooper DL, Krause DS . A preclinical xenotransplantation animal model to assess human hematopoietic stem cell engraftment. Transfusion 2004; 44: 555–566.

    Article  Google Scholar 

  9. Perez LE, Rinder HM, Wang C, Tracey JB, Maun N, Krause DS . Xenotransplantation of immunodeficient mice with mobilized human blood CD34+ cells provides an in vivo model for human megakaryocytopoiesis and platelet production. Blood 2001; 97: 1635–1643.

    Article  CAS  Google Scholar 

  10. Bruno S, Gunetti M, Gammaitoni L, Dane A, Cavalloni G, Sanavio F et al. In vitro and in vivo megakaryocyte differentiation of fresh and ex-vivo expanded cord blood cells: rapid and transient megakaryocyte reconstitution. Haematologica 2003; 88: 379–387.

    Google Scholar 

  11. Bruno S, Gunetti M, Gammaitoni L, Perissinotto E, Caione L, Sanavio F et al. Fast but durable megakaryocyte repopulation and platelet production in NOD/SCID mice transplanted with ex-vivo expanded human cord blood CD34+ cells. Stem Cells 2004; 22: 135–143.

    Article  Google Scholar 

  12. Feng Y, Zhang L, Xiao ZJ, Li B, Liu B, Fan CG et al. An effective and simple expansion system for megakaryocyte progenitor cells using a combination of heparin with thrombopoietin and interleukin-11. Exp Hematol 2005; 33: 1537–1543.

    Article  CAS  Google Scholar 

  13. van Hensbergen Y, Schipper LF, Brand A, Slot MC, Welling M, Nauta AJ et al. Ex vivo culture of human CD34+ cord blood cells with thrombopoietin (TPO) accelerates platelet engraftment in a NOD/SCID mouse model. Exp Hematol 2006; 34: 943–950.

    Article  CAS  Google Scholar 

  14. Schipper LF, Brand A, Reniers N, Melief CJ, Willemze R, Fibbe WE . Differential maturation of megakaryocyte progenitor cells from cord blood and mobilized peripheral blood. Exp Hematol 2003; 31: 324–330.

    Article  Google Scholar 

  15. van den Oudenrijn S, von dem Borne AE, de Haas M . Differences in megakaryocyte expansion potential between CD34(+) stem cells derived from cord blood, peripheral blood, and bone marrow from adults and children. Exp Hematol 2000; 28: 1054–1061.

    Article  CAS  Google Scholar 

  16. Dorrell C, Gan OI, Pereira DS, Hawley RG, Dick JE . Expansion of human cord blood CD34(+)CD38(−) cells in ex vivo culture during retroviral transduction without a corresponding increase in SCID repopulating cell (SRC) frequency: dissociation of SRC phenotype and function. Blood 2000; 95: 102–110.

    CAS  Google Scholar 

Download references

Acknowledgements

We thank our collaborators from the animal facilities. The stem cell laboratory at Sanquin Amsterdam is acknowledged for providing the peripheral blood progenitor cell samples and technical assistance. Professor D Roos and Dr PL Hordijk are acknowledged for critical reading of the article.

Author information

Authors and Affiliations

  1. Department of Experimental Immunohematology, Sanquin Research, Amsterdam, and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands

    M R Tijssen, F di Summa, J J Zwaginga, C E van der Schoot & C Voermans

  2. Department of Molecular Cell Biology, Sanquin Research, Amsterdam, and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands

    P B van Hennik

  3. Department of Immunohematology-Bloodtransfusion, Leiden University Medical Center, Leiden, The Netherlands

    J J Zwaginga

  4. Department of Hematology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands

    C E van der Schoot

Authors
  1. M R Tijssen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. P B van Hennik
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. F di Summa
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. J J Zwaginga
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. C E van der Schoot
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. C Voermans
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to C Voermans.

Additional information

Supplementary Information accompanies the paper on the Leukemia website (http://www.nature.com/leu)

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tijssen, M., van Hennik, P., di Summa, F. et al. Transplantation of human peripheral blood CD34-positive cells in combination with ex vivo generated megakaryocytes results in fast platelet formation in NOD/SCID mice. Leukemia 22, 203–208 (2008). https://doi.org/10.1038/sj.leu.2404979

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.leu.2404979

Search

Advanced search

Quick links