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. 1992 May 1;89(9):4013–4017. doi: 10.1073/pnas.89.9.4013

Growth factor requirements for survival in G0 and entry into the cell cycle of primitive human hemopoietic progenitors.

A G Leary 1, H Q Zeng 1, S C Clark 1, M Ogawa 1
PMCID: PMC525622  PMID: 1373892

Abstract

In this study we have isolated populations of dormant human hemopoietic progenitors by two different approaches. First CD34+ cells isolated by panning were further separated on the basis of absence of HLA-DR expression by using fluorescence-activated cell sorting. Second, CD34+ HLA-DR- cells were isolated by nonadherence to soybean agglutinin, negative immunomagnetic bead selection with lineage-specific antibodies, and two-color cell sorting. Progenitors in either cell population were unable to form colonies in the presence of interleukin (IL)-3 alone but yielded a substantial number of colonies, including multilineage colonies, in the presence of combinations of IL-3 and IL-6. Similarly, IL-3 plus any one of the other synergistic factors, including granulocyte colony-stimulating factor, IL-11, leukemia inhibitory factor, and steel factor, effectively supported colony formation from CD34+ HLA-DR- progenitors. Sequential observation of colony formation from single CD34+ HLA-DR- cells provided definitive evidence that the synergistic factors trigger cell divisions of dormant cells. Studies with delayed addition of factors to the cultures provided evidence that this population of cells also requires IL-3 or granulocyte/macrophage colony-stimulating factor (GM-CSF) to survive even while dormant. In contrast, none of the synergistic factors were able to replace IL-3 or GM-CSF in this function. These findings confirm and extend the model that multiple factors with overlapping functions operate both independently and in combination to regulate early stages of hemopoiesis.

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Selected References

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  1. Andrews R. G., Singer J. W., Bernstein I. D. Precursors of colony-forming cells in humans can be distinguished from colony-forming cells by expression of the CD33 and CD34 antigens and light scatter properties. J Exp Med. 1989 May 1;169(5):1721–1731. doi: 10.1084/jem.169.5.1721. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brandt J., Baird N., Lu L., Srour E., Hoffman R. Characterization of a human hematopoietic progenitor cell capable of forming blast cell containing colonies in vitro. J Clin Invest. 1988 Sep;82(3):1017–1027. doi: 10.1172/JCI113658. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Huang E., Nocka K., Beier D. R., Chu T. Y., Buck J., Lahm H. W., Wellner D., Leder P., Besmer P. The hematopoietic growth factor KL is encoded by the Sl locus and is the ligand of the c-kit receptor, the gene product of the W locus. Cell. 1990 Oct 5;63(1):225–233. doi: 10.1016/0092-8674(90)90303-v. [DOI] [PubMed] [Google Scholar]
  4. Ikebuchi K., Clark S. C., Ihle J. N., Souza L. M., Ogawa M. Granulocyte colony-stimulating factor enhances interleukin 3-dependent proliferation of multipotential hemopoietic progenitors. Proc Natl Acad Sci U S A. 1988 May;85(10):3445–3449. doi: 10.1073/pnas.85.10.3445. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Ikebuchi K., Wong G. G., Clark S. C., Ihle J. N., Hirai Y., Ogawa M. Interleukin 6 enhancement of interleukin 3-dependent proliferation of multipotential hemopoietic progenitors. Proc Natl Acad Sci U S A. 1987 Dec;84(24):9035–9039. doi: 10.1073/pnas.84.24.9035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Iscove N. N., Sieber F., Winterhalter K. H. Erythroid colony formation in cultures of mouse and human bone marrow: analysis of the requirement for erythropoietin by gel filtration and affinity chromatography on agarose-concanavalin A. J Cell Physiol. 1974 Apr;83(2):309–320. doi: 10.1002/jcp.1040830218. [DOI] [PubMed] [Google Scholar]
  7. Leary A. G., Hirai Y., Kishimoto T., Clark S. C., Ogawa M. Survival of hemopoietic progenitors in the G0 period of the cell cycle does not require early hemopoietic regulators. Proc Natl Acad Sci U S A. 1989 Jun;86(12):4535–4538. doi: 10.1073/pnas.86.12.4535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Leary A. G., Ikebuchi K., Hirai Y., Wong G. G., Yang Y. C., Clark S. C., Ogawa M. Synergism between interleukin-6 and interleukin-3 in supporting proliferation of human hematopoietic stem cells: comparison with interleukin-1 alpha. Blood. 1988 Jun;71(6):1759–1763. [PubMed] [Google Scholar]
  9. Leary A. G., Ogawa M. Blast cell colony assay for umbilical cord blood and adult bone marrow progenitors. Blood. 1987 Mar;69(3):953–956. [PubMed] [Google Scholar]
  10. Leary A. G., Wong G. G., Clark S. C., Smith A. G., Ogawa M. Leukemia inhibitory factor differentiation-inhibiting activity/human interleukin for DA cells augments proliferation of human hematopoietic stem cells. Blood. 1990 May 15;75(10):1960–1964. [PubMed] [Google Scholar]
  11. Leary A. G., Yang Y. C., Clark S. C., Gasson J. C., Golde D. W., Ogawa M. Recombinant gibbon interleukin 3 supports formation of human multilineage colonies and blast cell colonies in culture: comparison with recombinant human granulocyte-macrophage colony-stimulating factor. Blood. 1987 Nov;70(5):1343–1348. [PubMed] [Google Scholar]
  12. Lu L., Walker D., Broxmeyer H. E., Hoffman R., Hu W., Walker E. Characterization of adult human marrow hematopoietic progenitors highly enriched by two-color cell sorting with My10 and major histocompatibility class II monoclonal antibodies. J Immunol. 1987 Sep 15;139(6):1823–1829. [PubMed] [Google Scholar]
  13. Musashi M., Clark S. C., Sudo T., Urdal D. L., Ogawa M. Synergistic interactions between interleukin-11 and interleukin-4 in support of proliferation of primitive hematopoietic progenitors of mice. Blood. 1991 Sep 15;78(6):1448–1451. [PubMed] [Google Scholar]
  14. Musashi M., Yang Y. C., Paul S. R., Clark S. C., Sudo T., Ogawa M. Direct and synergistic effects of interleukin 11 on murine hemopoiesis in culture. Proc Natl Acad Sci U S A. 1991 Feb 1;88(3):765–769. doi: 10.1073/pnas.88.3.765. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Nakahata T., Ogawa M. Identification in culture of a class of hemopoietic colony-forming units with extensive capability to self-renew and generate multipotential hemopoietic colonies. Proc Natl Acad Sci U S A. 1982 Jun;79(12):3843–3847. doi: 10.1073/pnas.79.12.3843. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ogawa M. Effects of hemopoietic growth factors on stem cells in vitro. Hematol Oncol Clin North Am. 1989 Sep;3(3):453–464. [PubMed] [Google Scholar]
  17. Paul S. R., Bennett F., Calvetti J. A., Kelleher K., Wood C. R., O'Hara R. M., Jr, Leary A. C., Sibley B., Clark S. C., Williams D. A. Molecular cloning of a cDNA encoding interleukin 11, a stromal cell-derived lymphopoietic and hematopoietic cytokine. Proc Natl Acad Sci U S A. 1990 Oct;87(19):7512–7516. doi: 10.1073/pnas.87.19.7512. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Suda T., Suda J., Ogawa M., Ihle J. N. Permissive role of interleukin 3 (IL-3) in proliferation and differentiation of multipotential hemopoietic progenitors in culture. J Cell Physiol. 1985 Aug;124(2):182–190. doi: 10.1002/jcp.1041240203. [DOI] [PubMed] [Google Scholar]
  19. Suda T., Suda J., Ogawa M. Proliferative kinetics and differentiation of murine blast cell colonies in culture: evidence for variable G0 periods and constant doubling rates of early pluripotent hemopoietic progenitors. J Cell Physiol. 1983 Dec;117(3):308–318. doi: 10.1002/jcp.1041170305. [DOI] [PubMed] [Google Scholar]
  20. Terstappen L. W., Huang S., Safford M., Lansdorp P. M., Loken M. R. Sequential generations of hematopoietic colonies derived from single nonlineage-committed CD34+CD38- progenitor cells. Blood. 1991 Mar 15;77(6):1218–1227. [PubMed] [Google Scholar]
  21. Williams D. E., Eisenman J., Baird A., Rauch C., Van Ness K., March C. J., Park L. S., Martin U., Mochizuki D. Y., Boswell H. S. Identification of a ligand for the c-kit proto-oncogene. Cell. 1990 Oct 5;63(1):167–174. doi: 10.1016/0092-8674(90)90297-r. [DOI] [PubMed] [Google Scholar]
  22. Zsebo K. M., Wypych J., McNiece I. K., Lu H. S., Smith K. A., Karkare S. B., Sachdev R. K., Yuschenkoff V. N., Birkett N. C., Williams L. R. Identification, purification, and biological characterization of hematopoietic stem cell factor from buffalo rat liver--conditioned medium. Cell. 1990 Oct 5;63(1):195–201. doi: 10.1016/0092-8674(90)90300-4. [DOI] [PubMed] [Google Scholar]

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