Stimulation of Human Early and Late Erythropoietic Progenitor Cells by Insulin: Evidence for Different Mechanisms

  • Gunther Konwalinka
  • Christian J. Wiedermann
  • Andreas Petzer
  • Kurt Grunewald
  • Christoph Breier
  • Josef Patsch
  • Dietmar Geissler
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 34)


In order to investigate cellular mechanisms involved in insulin stimulation of erythropoiesis, we have studied the response of early (BFU-e) and late (CFU-e) erythroid progenitor cells in a serum-free agar culture system. In this assay system, CFU-e proliferation occurred in media containing low-density lipoproteins, bovine serum albumin, transferrin and recombinant erythropoietin (rEPO). Insulin in physiological concentrations as low as 10-12M, added directly to cultures, augmented CFU-e colony formation. This stimulatory effect was also seen when monocyte- and T lymphocyte-depleted cells from normal donors were cultured. In contrast, BFU-e was not stimulated by media devoid of insulin. Occurrence of BFU-e colonies required the presence of insulin in concentrations higher than 10-8M This insulin effect was not dependent on the presence of monocytes and T lymphocytes. Delayed addition studies of rEPO to insulin containing cultures revealed a slight but significant survival rate of CFU-e. A similar survival rate was found for BFU-e. From this, we conclude that insulin stimulates CFU-e by an EPO-like activity. For BFU-e, however, the decline in the number of bursts caused by EPO deprivation implies that insulin does not act directly as a burst-promoting activity but that it probably induces the release of this activity from non-adherent and T lymphocyte-depleted bone marrow cells.


Colony Formation Hemopoietic Stem Cell Recombinant Erythropoietin Erythroid Progenitor Cell Normal Human Bone Marrow 


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  1. 1.
    Yokota, T., F. Lee, D. Rennick, C. Hall, N. Arai, T. Mosmann, G. Nabel, H. Cantor, and K. Arai. 1984. Isolation and characterization of a mouse cDNA clone that expresses mast-cell growth-factor activity in monkey cells. Proc. Natl. Acad. Sci. USA 81: 1070–1074.PubMedCrossRefGoogle Scholar
  2. 2.
    Jacobs, K., C. Shoemaker, R. Rudersdorf, S.D. Neill, R.J. Kaufmann, A. Mufson, J. Seehra, S.S. Jones, R. Hewick, E.F. Fritsch, M. Kawakita, T. Shimizu, and T. Miyake. 1985. Isolation and characterization of genomic and cDNA clones of human erythropoietin. Nature 313: 806–810.PubMedCrossRefGoogle Scholar
  3. 3.
    Lin, F., S. Suggs, C. Lin, J.K. Browne, R. Smallins, J.C. Egrie, K.K. Chen, G.M. Fox, F. Martin, Z. Stabinsky, S.M. Badrawi, P. Lai and E. Goldwasser. 1985. Cloning and expression of the human erythropoietin gene. Proc. Natl. Acad. Sci. USA 82: 7580–7584.PubMedCrossRefGoogle Scholar
  4. 4.
    Golde, D.W. 1978. Hormonal modulation of erythropoiesis in vitro. In: In Vitro Aspects of Erythropoiesis. M.J. Murphy Jr., C. Peschle, A.S. Gordon, and E.A. Mirand, editors. Springer Verlag, New York. 81–85.CrossRefGoogle Scholar
  5. 5.
    Dainiak, N. 1985. Role of defined and undefined serum additives to hemopoietic stem cell culture. In Hemopoietic Stem Cell Physiology. E.P. Cronkite, N. Dainiak, R.P. McCaffery, J. Palek, P.J. Quesenbery, editors. Liss, New York. 59–68.Google Scholar
  6. 6.
    Bersch, N., J.E. Groopman and D. W. Golde. 1982. Natural and biosynthetic insulin stimulates the growth of human erythroid progenitors in vitro. J. Clin. Endocrinol. Metab. 55: 1209–1211.PubMedCrossRefGoogle Scholar
  7. 7.
    Kurtz, A., W. Jelkmann and C. Bauer. 1983. Insulin stimulates erythroid colony formation independently of erythropoietin. Br. J. Haematol. 53: 311–316.PubMedCrossRefGoogle Scholar
  8. 8.
    Dainiak, N., and S. Kreczko. 1985. Interactions of insulin, insulinlike growth factor II, and platelet-derived growth factor in erythropoietic culture. J. Clin. Invest. 76: 1237–1242.PubMedCrossRefGoogle Scholar
  9. 9.
    Geffner, M.E., S.A. Kaplan, N. Bersch, B.M. Lippe, W.G. Smith, R.A. Nagel, T.V. Santulli Jr,, C.H. Li, and D.W. Golde. 1987. Leprechaunism: In vitro insulin action despite genetic insulin resistance. Ped. Res. 22: 286–291.CrossRefGoogle Scholar
  10. 10.
    Flier, J.S., P. Usher, and A.C. Moses. 1986. Monoclonal antibody to the type I insulin-like growth factor (igF-l) receptor blocks IgF-I receptor-mediated DNA synthesis: Clarification of the mitogenic mechanisms of IgF-I and insulin in human skin fibroblasts. Proc. Natl. Acad. Sci. USA 83: 664–668.PubMedCrossRefGoogle Scholar
  11. 11.
    Chen, P., S. Kwan, T. Hwang, B.J. Chiang and C. Chou. 1983. Insulin receptors on leukemia and lymphoma cells. Blood 62: 251–255.PubMedGoogle Scholar
  12. 12.
    Konwalinka, G., C. Breier, D. Geissler, C. Peschel, C.J. Wiedermann, J. Patsch and H. Braunsteiner. 1988. Proliferation and Differentiation of Human Erythropoiesis in Vitro: Effect of Different Human Lipoprotein Species. Exp. Hematol. 16: 125–130.PubMedGoogle Scholar
  13. 13.
    Iscove, N.N., L.J. Guilbert, and C. Weymann. 1980. Complete replacement of serum in primary culture of erythropoietin-dependent red cell precursors (CFU-e) by albumin, transferrin, iron, unsaturated fatty acid, lecithin and cholesterol. Exp. Cell Res. 126: 121–131.PubMedCrossRefGoogle Scholar
  14. 14.
    Havel, R.J., H.A. Eder, and J.H. Bragdon. 1955. Distribution and chemical composition of ultra-centrifugally separated lipopro-teins in human serum. J. Clin. Invest. 34: 1345–1351.PubMedCrossRefGoogle Scholar
  15. 15.
    Konwalinka, G., D. Geissler, C. Peschel, C. Breier, K. Grunewald, R. Odavic and H. Braunsteiner. 1986. Human erythropoiesis in vitro and the source of burst-promoting activity in a serum-free system. Exp. Hematol. 14: 899–903.PubMedGoogle Scholar
  16. 16.
    Konwalinka, G., C. Peschel, J. Boyd, D. Geissler, M. Ogriseg, R. Odavic and H. Braunsteiner. 1984. A miniaturized agar culture system for cloning human erythropoietic progenitor cells. Exp. Hematol. 12: 75–79.PubMedGoogle Scholar
  17. 17.
    Schroder, K.S., S. Raptis, W. Beischer. 1976. Radio-immunologische Bestimmunung von STH, Insulin und C-Peptid. Med. Welt. 27: 1720–1726.PubMedGoogle Scholar
  18. 18.
    Davis, R.J., M. Faucher, L.K. Racaniello, A. Carruthers and M.P. Czech. 1987. Insulin-like growth factor I and epidermal growth factor regulate the expression of transferrin receptors at the cell surface by distinct mechanisms. J. Biol. Chem. 262: 13126–13135.PubMedGoogle Scholar
  19. 19.
    Dainiak, N., D. Sutter, and S. Kreczko. 1986. L-tri-iodothyronine augments erythropoietic growth factor release from peripheral blood and bone marrow leukocytes. Blood 68: 1289–1297.PubMedGoogle Scholar
  20. 20.
    Zuckerman, K.S. 1981. Human Erythroid burst-forming units. Growth in vitro is dependent on monocytes but not T lymphocytes. J. Clin. Invest. 67: 702–710.PubMedCrossRefGoogle Scholar
  21. 21.
    Kirschner, J., H. Ozer, M. O’Leary, D. Highyand, M. Marinello, and H. Preisler. 1985. T cell depletion of bone marrow does not inhibit in vitro hematopoiesis. Blut. 50: 201–206.CrossRefGoogle Scholar
  22. 22.
    Dainiak, N., A. Najman, V. Kulkarni, D. Sutter, S. Kreczko, L. Feldman, C. Baillou, and G. Leblanc. 1984. Regulation of erythroid burst proliferation by B lymphocytes from healthy donors and patients with chronic lymphocytic leukemia (CLL). Blood 64: 113–118. (Suppl.)Google Scholar
  23. 23.
    Iscove, N.N. and L.J. Guilbert. 1978. Erythropoietin-independence of early erythropoiesis and a two-regulator model of proliferative control in the hemopoietic system. In Vitro Aspects of Erythro-poiesis. M. J. Murphy. Springer Verlag, New York. 3–7.CrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1988

Authors and Affiliations

  • Gunther Konwalinka
    • 1
  • Christian J. Wiedermann
    • 1
  • Andreas Petzer
    • 1
  • Kurt Grunewald
    • 1
  • Christoph Breier
    • 1
  • Josef Patsch
    • 1
  • Dietmar Geissler
    • 1
  1. 1.Department of Internal MedicineUniversity of InnsbruckInnsbruckAustria

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