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Thrombopoietin Supports the Continuous Growth of Cytokine-Dependent Human Leukemia Cell Lines

  • H. G. Drexler
  • M. Zaborski
  • H. Quentmeier
Conference paper
Part of the Haematology and Blood Transfusion / Hämatologie und Bluttransfusion book series (HAEMATOLOGY, volume 39)

Abstract

The search for a factor regulating megakaryocytopoiesis lead to the cloning of thrombopoietin (TPO) that specifically promotes proliferation and differentiation of the megakaryocytic lineage. The availability of recombinant TPO and its imminent clinical use has made a more detailed understanding of its effects on hematopoietic cells more urgent. Normal megakaryocyto- and thrombopoiesis occurs predominantly in the bone marrow, a difficult organ to study in situ, particular in humans due to the low numbers of megakaryocytic progenitors and the difficult isolation as pure populations.We developed an in vitro system which may allow to address questions regarding the biology of TPO. The acute myeloid leukemia (AML)-derived cell lines HU-3, M-07e, M-MOK and TF-1 have absolute dependence on granulocyte-macrophage colony-stimulating factor (GM-CSF). We cultured these cells long term (> 6 months) in the presence of TPO (omitting GM-CSF). TPO alone supported the maintenance and expansion of these sister cell lines, HU-3/TPO, M-07e/TPO, M-MOK/TPO and TF-1/TPO, that displayed longer doubling times, a larger cell size, and a higher percentage of poly-nucleated giant cells and slightly adherent cells than the corresponding countercultures grown with GM-CSF. In the absence of TPO, the cells died quickly within few days; thus, the TPO-grown cell lines have an absolute dependence on this factor, but could all be switched back to growth with GM-CSF. In comparison with the GM-CSF-treated cells, the receptors for GM-CSF and interleukin-3 (IL-3) were down-regulated and the receptors for stem cell factor (SCF) and TPO were up-regulated in the TPO-exposed cells. A short-term proliferation assay showed a stronger response of the TPO-cell lines to erythropoietin, GM-CSF, IL-3, PIXY-321, SCF and TPO than the GM-CSF-cell lines. Flow cytometric analysis of the GM-CSFand TPO-cultured lines displayed an up-regulation of the megakaryocytic surface markers CD41, CD42 and CD61 and a down-regulation of the erythroid marker glycophorin A in the latter cell lines, suggesting differentiation along the megakaryocytic lineage. Thus, in long term exposure, TPO appears to have both a proliferative and a differentiative effect on responsive cells. Under serum-deprived culture conditions, TPO acted as a survival factor on the TPO-cell lines. Taken together, these findings indicate that the TPO-dependent cell lines represent important biological reagents for further characterization of the biology of TPO and should provide also a great aid for future in vitro experiments aimed at elucidating megakaryocyto- and thrombopoiesis.

Keywords

Acute Myeloid Leukemia Conditioned Medium Stem Cell Factor Leukemia Cell Line Bladder Carcinoma Cell Line 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. 1.
    Drexler HG, Quentmeier H (1996) Thrombopoietin: Expression of its receptor MPL and proliferative effects on leukemic cells. Leukemia 10: 1405–1421PubMedGoogle Scholar
  2. 2.
    Kaushansky K (1996) Thrombopoietin: Biological and preclinical properties. Leukemia 10, Suppl 1: S46 - S48Google Scholar
  3. 3.
    Quentmeier H, Zaborski M, Graf G, Ludwig WD, Drexler HG (1996) Expression of the receptor MPL and proliferative effects of its ligand thrombopoietin on human leukemia cell lines. Leukemia 10: 297–310PubMedGoogle Scholar
  4. 4.
    Drexler HG, Gignac SM, Minowada J (1994) Hematopoietic cell lines. In: Hay RJ, Park JG, Gaz-dar A (eds) Atlas of human tumor cell lines. Academic Press, Orlando, pp 213–250Google Scholar
  5. 5.
    Drexler HG, Dirks W, MacLeod RAF, Quentmeier H, Steube K (eds) (1995) DSM Catalogue of human and animal cell lines, 5th edn, Braunschweig, GermanyGoogle Scholar
  6. 6.
    Morgan D, Class R, Soslau G, Brodsky I (1996) Cytokine-mediated erythroid maturation in CD4+ megakaryoblastic human cell line HU-3. Exp Hematol, submittedGoogle Scholar
  7. 7.
    Avanzi GC, Lista P, Giovinazzo B, Miniero R, Saglio G, Benetton G, Coda R, Cattoretti G, Pegoraro L (1988) Selective growth response to IL-3 of a human leukaemic cell line with megakaryoblastic features. Brit J Haematol 69: 359–366CrossRefGoogle Scholar
  8. 8.
    Morgan DA, Gumucio DL, Brodsky I (1991) Granulocyte-macrophage colony-stimulating factordependent growth and erythropoietin-induced differentiation of a human cell line MB-02. Blood 78: 2860–2871PubMedGoogle Scholar
  9. 9.
    Itano M, Tsuchiya S, Minegishi N, Fujie H, Minegishi M, Morita S, Yambe T, Ohashi Y, Masuda T, Koike T, Konno T (1995) Establishment and characterization of a novel human immature megakaryoblastic leukemia cell line, M-MOK, dependent on fibroblasts for its viability. Exp Hematol 23: 1301–1309Google Scholar
  10. 10.
    Hu ZB, Ma W, Zaborski M, MacLeod RAF, Quentmeier H, Drexler HG (1996) Establishment and characterization of two novel cytokine-responsive acute myeloid and monocytic leukemia cell lines, MUTZ-2 and MUTZ-3. Leukemia 10: 1025–1040PubMedGoogle Scholar
  11. 11.
    Wang C, Koistinen P, Yang GS, Williams DE, Lyman SD, Minden MD, McCulloch EA (1991) Mast cell growth factor, a ligand for the receptor encoded by c-kit, affects the growth in culture of the blast cells of acute myeloblastic leukemia. Leukemia 5: 493–499PubMedGoogle Scholar
  12. 12.
    Kitamura T, Tojo A, Kuwaki T, Chiba S, Miyazono K, Urabe A, Takaku F (1989) Identification and analysis of human erythropoietin receptors on a factor-dependent cell line, TF-1. Blood 73: 375–380PubMedGoogle Scholar
  13. 13.
    Komatsu N, Nakauchi H, Miwa A, Ishihara T,Eguchi M, Moroi M, Okada M, Sato Y, Wada H, Yawata Y, Suda T, Miura Y (1991) Establishment and characterization of a human leukemic cell line with megakaryocytic features: Dependency on granulocyte-macrophage colony-stimulating factor, interleukin 3, or erythropoietin for growth and survival. Cancer Res 51: 341–348PubMedGoogle Scholar
  14. 13.
    Komatsu N, Nakauchi H, Miwa A, Ishihara T,Eguchi M, Moroi M, Okada M, Sato Y, Wada H, Yawata Y, Suda T, Miura Y (1991) Establishment and characterization of a human leukemic cell line with megakaryocytic features: Dependency on granulocyte-macrophage colony-stimulating factor, interleukin 3, or erythropoietin for growth and survival. Cancer Res 51: 341–348PubMedGoogle Scholar
  15. 15.
    Komatsu N, Kunitama M, Yamada M, Hagiwara T, Kato T, Miyazaki H, Eguchi M, Yamamoto M, Miura Y (1996) Establishment and characterization of the thrombopoietin-dependent megakaryo-cytic cell line, UT-7/TPO. Blood 87: 4552–4560PubMedGoogle Scholar
  16. 16.
    Komatsu N, Yamamoto M, Fujita H, Miwa A, Ha-take K, Endo T, Okano H, Katsube T, Fukumaki Y, Sassa S, Miuta Y (1993) Establishment and characterization of an erythropoietin-dependent subline, UT-7/Epo, derived from human leukemia cell line, UT-7. Blood 82: 456–464PubMedGoogle Scholar
  17. 17.
    Minegishi N, Minegishi M, Tsuchiya S, Fujie H, Nagai T, Hayashi N, Yamamoto M, Konnop T (1994) Erythropoietin-dependent induction of hemoglobin synthesis in a cytokine-dependent cell line M-TAT. J Biol Chem 269: 27700–27704PubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1998

Authors and Affiliations

  • H. G. Drexler
    • 1
  • M. Zaborski
    • 1
  • H. Quentmeier
    • 1
  1. 1.Department of Human Animal Cell CulturesDSMZ-German Collection of Microorganisms and Cell CulturesBraunschweigGermany

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