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Stem Cell Expansion

Platelet-derived growth factor enhances ex vivo expansion of megakaryocytic progenitors from human cord blood

Bone Marrow Transplantation volume 27, pages 1075–1080 (2001)Cite this article

Abstract

Infusion of ex vivo expanded megakaryocytic (MK) progenitor cells is a strategy for shortening the duration of thrombocytopenia after haematopoietic stem cell transplantation. The cell dose after expansion has emerged as a critical factor for achieving the desired clinical outcomes. This study aimed to establish efficient conditions for the expansion of the MK lineage from enriched CD34+ cells of umbilical cord blood and to investigate the effect of platelet-derived growth factor (PDGF) in this system. Our results demonstrated that thrombopoietin (TPO) alone produced a high proportion of CD61+CD41+ cells but a low total cell count and high cell death, resulting in an inferior expansion. The addition of interleukin-1β (IL-1β), Flt-3 ligand (Flt-3L) and to a lesser extent IL-3 improved the expansion outcome. The treatment groups with three to five cytokines produced efficient expansions of CFU-MK up to 400-fold with the highest yield observed in the presence of TPO, IL-1β, IL-3, IL-6 and Flt-3L. CD34+ cells were expanded by five to 22-fold. PDGF improved the expansion of all cell types with CD61+CD41+ cells, CFU-MK and CD34+ cells increased by 101%, 134% and 70%, respectively. On day 14, the CD61+ population consisted of diploid (86.5%), tetraploid (11.8%) and polyploid (8N–32N; 1.69%) cells. Their levels were not affected by PDGF. TPO, IL-1β, IL-3, IL-6, Flt-3L and PDGF represented an effective cytokine combination for expanding MK progenitors while maintaining a moderate increase of CD34+ cells. This study showed, for the first time, that PDGF enhanced the ex vivo expansion of the MK lineage, without promoting their in vitro maturation. PDGF might be a suitable growth factor to improve the ex vivo expansion of MK progenitors for clinical applications. Bone Marrow Transplantation (2001) 27, 1075–1080.

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References

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

    Article  CAS  Google Scholar 

  2. Gluckman E, Locatelli F . Umbilical cord blood transplants Curr Opin Hematol 2000 7: 353–357

    Article  CAS  Google Scholar 

  3. Bertolini F, Battaglia M, Pedrazzoli P 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  PubMed  Google Scholar 

  4. Paquette RL, Dergham ST, Karpf E et al. Ex vivo expanded unselected peripheral blood: progenitor cells reduce posttransplantation neutropenia, thrombocytopenia, and anemia in patients with breast cancer Blood 2000 96: 2385–2390

    CAS  PubMed  Google Scholar 

  5. Stiff P, Chen B, Franklin W et al. Autologous transplantation of ex vivo expanded bone marrow cells grown from small aliquots after high-dose chemotherapy for breast cancer Blood 2000 95: 2169–2174

    CAS  PubMed  Google Scholar 

  6. Guerriero R, Testa U, Gabbianelli M et al. Unilineage megakaryocytic proliferation and differentiation of purified hematopoietic progenitors in serum-free liquid culture Blood 1995 86: 3725–3736

    CAS  PubMed  Google Scholar 

  7. Debili N, Wendling F, Katz A et al. The Mpl-ligand or thrombopoietin or megakaryocyte growth and differentiative factor has both direct proliferative and differentiative activities on human megakaryocyte progenitors Blood 1995 86: 2516–2525

    CAS  PubMed  Google Scholar 

  8. Williams JL, Pipia GG, Datta NS, Long MW . Thrombopoietin requires additional megakaryocyte-active cytokines for optimal ex vivo expansion of megakaryocyte precursor cells Blood 1998 91: 4118–4126

    CAS  Google Scholar 

  9. Yang M, Li K, Chui CM et al. Expression of interleukin (IL) 1 type I and type II receptors in megakaryocytic cells and enhancing effects of IL-1beta on megakaryocytopoiesis and NF-E2 expression Br J Haematol 2000 111: 371–380

    Article  CAS  Google Scholar 

  10. Dolzhanskiy A, Basch RS, Karpatkin S . The development of human megakaryocytes: III. Development of mature megakaryocytes from highly purified committed progenitors in synthetic culture media and inhibition of thrombopoietin-induced polyploidization by interleukin-3 Blood 1997 89: 426–434

    CAS  PubMed  Google Scholar 

  11. Catani L, Gugliotta L, Campanini E et al. Megakaryocyte progenitors derived from bone marrow or G-CSF-mobilized peripheral blood CD34 cells show a distinct phenotype and responsiveness to interleukin-3 (IL-3) and PEG-recombinant human megakaryocyte growth and development factor (PEG-rHuMGDF) Br J Haematol 1998 100: 207–218

    Article  CAS  Google Scholar 

  12. Navarro S, Debili N, Le Couedic JP et al. Interleukin-6 and its receptor are expressed by human megakaryocytes: in vitro effects on proliferation and endoreplication Blood 1991 77: 461–471

    CAS  PubMed  Google Scholar 

  13. Teramura M, Kobayashi S, Hoshino S et al. Interleukin-11 enhances human megakaryocytopoiesis in vitro Blood 1992 79: 327–331

    CAS  PubMed  Google Scholar 

  14. Li K, Yang M, Lam AC et al. Effects of flt-3 ligand in combination with TPO on the expansion of megakaryocytic progenitors Cell Transplant 2000 9: 125–131

    Article  CAS  Google Scholar 

  15. Briddell RA, Bruno E, Cooper RJ et al. Effect of c-kit ligand on in vitro human megakaryocytopoiesis Blood 1991 78: 2854–2859

    CAS  PubMed  Google Scholar 

  16. Heldin CH, Westermark B, Wasteson A . Platelet-derived growth factor: purification and partial characterization Proc Natl Acad Sci USA 1979 76: 3722–3726

    Article  CAS  Google Scholar 

  17. Deuel TF, Huang JS, Proffitt RT et al. Human platelet-derived growth factor. Purification and resolution into two active protein fractions J Biol Chem 1981 256: 8896–8899

    CAS  PubMed  Google Scholar 

  18. Khachigian LM, Chesterman CN . Platelet-derived growth factor and alternative splicing: a review Pathology 1992 24: 280–290

    Article  CAS  Google Scholar 

  19. Westermark B, Claesson-Welsh L, Heldin CH . Structural and functional aspects of the receptors for platelet-derived growth factor Prog Growth Factor Res 1989 1: 253–266

    Article  CAS  Google Scholar 

  20. Khachigian LM, Chesterman CN . Platelet-derived growth factor and alternative splicing: a review Pathology 1992 24: 280–290

    Article  CAS  Google Scholar 

  21. Leveen P, Pekny M, Gebre-Medhin S et al. Mice deficient for PDGF B show renal, cardiovascular, and hematological abnormalities Genes Dev 1994 8: 1875–1887

    Article  CAS  Google Scholar 

  22. Soriano P . Abnormal kidney development and hematological disorders in PDGF beta-receptor mutant mice Genes Dev 1994 8: 1888–1896

    Article  CAS  Google Scholar 

  23. Dainiak N, Davies G, Kalmanti M et al. Platelet-derived growth factor promotes proliferation of erythropoietic progenitor cells in vitro J Clin Invest 1983 71: 1206–1214

    Article  CAS  Google Scholar 

  24. Michalevicz R, Katz F, Stroobant P et al. Platelet-derived growth factor stimulates growth of highly enriched multipotent haemopoietic progenitors Br J Haematol 1986 63: 591–598

    Article  CAS  Google Scholar 

  25. Yang M, Khachigian LM, Hicks C et al. Identification of PDGF receptors on human megakaryocytes and megakaryocytic cell lines Thromb Haemost 1997 78: 892–896

    Article  CAS  Google Scholar 

  26. Yang M, Chesterman CN, Chong BH . Recombinant PDGF enhances megakaryocytopoiesis in vitro Br J Haematol 1995 91: 285–289

    Article  CAS  Google Scholar 

  27. Liu J, Li K, Yuen PM et al. Ex vivo expansion of enriched CD34+ cells from neonatal blood in the presence of thrombopoietin, a comparison with cord blood and bone marrow Bone Marrow Transplant 1999 24: 247–252

    Article  CAS  Google Scholar 

  28. Kogler G, Nurnberger W, Fischer J et al. Simultaneous cord blood transplantation of ex vivo expanded together with non-expanded cells for high risk leukemia Bone Marrow Transplant 1999 24: 397–403

    Article  CAS  Google Scholar 

  29. Pecora AL, Stiff P, Jennis A et al. Prompt and durable engraftment in two older adult patients with high risk chronic myelogenous leukemia (CML) using ex vivo expanded and unmanipulated unrelated umbilical cord blood Bone Marrow Transplant 2000 25: 797–799

    Article  CAS  Google Scholar 

  30. McNiece I, Jones R, Bearman SI et al. Ex vivo expanded peripheral blood progenitor cells provide rapid neutrophil recovery after high-dose chemotherapy in patients with breast cancer Blood 2000 96: 3001–3007

    CAS  PubMed  Google Scholar 

  31. van den Oudenrijn S, de Haas M, Calafat J et al. 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 

  32. Veiby OP, Jacobsen FW, Cui L et al. The flt3 ligand promotes the survival of primitive hemopoietic progenitor cells with myeloid as well as B lymphoid potential. Suppression of apoptosis and counteraction by TNF-alpha and TGF-beta J Immunol 1996 157: 2953–2960

    CAS  PubMed  Google Scholar 

  33. Sungaran R, Chisholm OT, Markovic B et al. The role of platelet α-granular proteins in the regulation of thrombopoietin messenger RNA expression in human bone marrow stromal cells Blood 2000 95: 3094–3101

    CAS  PubMed  Google Scholar 

  34. Yang M, Li K, Lam AC et al. PDGF enhances granulo–monocytopoiesis via bone marrow stromal cells Int J Hematol 2001 73: 327–334

    Article  CAS  Google Scholar 

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Acknowledgements

We thank Ms Cecilia Mei Yan Chui and nurses of the Labor Ward for cord blood collection and the Industrial Support Fund AF/203/98 (Industry Department, the Government of the Hong Kong Special Administrative Region) for financial support.

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Authors and Affiliations

  1. Department of Paediatrics, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong

    RJ Su, K Li, M Yang, XB Zhang, TF Fok, CK Li & PMP Yuen

  2. Department of Anatomical and Cellular Pathology, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong

    KS Tsang

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Su, R., Li, K., Yang, M. et al. Platelet-derived growth factor enhances ex vivo expansion of megakaryocytic progenitors from human cord blood. Bone Marrow Transplant 27, 1075–1080 (2001). https://doi.org/10.1038/sj.bmt.1703042

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