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. 1986 Mar;77(3):894–899. doi: 10.1172/JCI112387

Declining sensitivity to interleukin 3 of murine multipotential hemopoietic progenitors during their development. Application to a culture system that favors blast cell colony formation.

K Koike, J N Ihle, M Ogawa
PMCID: PMC423476  PMID: 3081577

Abstract

When spleen cells of 5-fluorouracil (5-FU)-treated mice were cultured in the presence of interleukin 3 (IL-3), most colonies revealed IL-3 concentration-dependent colony formation except for mast cell colonies and blast cell colonies. While most colonies were smaller in lower concentrations of IL-3, the size of the blast cell colonies were similar between high and low IL-3 groups. These data suggested that blast cell colony development requires less IL-3 than the development of multilineage colonies from blast cell colonies. This notion was supported by experiments in which IL-3 was added twice to cultures of spleen cells of 5-FU-treated mice. When low concentrations of IL-3 were added on day 7, there was a reduction in the number of multilineage colonies formed without an effect on the number of blast cell colonies. Using this information, we developed a culture system that favors blast cell colony formation by cells of normal mice. When low (20 U/ml) concentrations of IL-3 were added to cultures of spleen cells of normal mice on day 7 of incubation in media containing 2-5% fetal calf serum, blast cell colonies were the predominant colony type. The blast cell colonies revealed high but variable secondary replating efficiencies. These data suggest that multipotential progenitors may become less sensitive to IL-3 as they differentiate in culture.

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Selected References

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  1. Beverley P. C., Linch D., Delia D. Isolation of human haematopoietic progenitor cells using monoclonal antibodies. Nature. 1980 Sep 25;287(5780):332–333. doi: 10.1038/287332a0. [DOI] [PubMed] [Google Scholar]
  2. Civin C. I., Strauss L. C., Brovall C., Fackler M. J., Schwartz J. F., Shaper J. H. Antigenic analysis of hematopoiesis. III. A hematopoietic progenitor cell surface antigen defined by a monoclonal antibody raised against KG-1a cells. J Immunol. 1984 Jul;133(1):157–165. [PubMed] [Google Scholar]
  3. Griffin J. D., Beveridge R. P., Schlossman S. F. Isolation of myeloid progenitor cells from peripheral blood of chronic myelogenous leukemia patients. Blood. 1982 Jul;60(1):30–37. [PubMed] [Google Scholar]
  4. Hodgson G. S., Bradley T. R. Properties of haematopoietic stem cells surviving 5-fluorouracil treatment: evidence for a pre-CFU-S cell? Nature. 1979 Oct 4;281(5730):381–382. doi: 10.1038/281381a0. [DOI] [PubMed] [Google Scholar]
  5. Ihle J. N., Keller J., Henderson L., Klein F., Palaszynski E. Procedures for the purification of interleukin 3 to homogeneity. J Immunol. 1982 Dec;129(6):2431–2436. [PubMed] [Google Scholar]
  6. Ihle J. N., Keller J., Oroszlan S., Henderson L. E., Copeland T. D., Fitch F., Prystowsky M. B., Goldwasser E., Schrader J. W., Palaszynski E. Biologic properties of homogeneous interleukin 3. I. Demonstration of WEHI-3 growth factor activity, mast cell growth factor activity, p cell-stimulating factor activity, colony-stimulating factor activity, and histamine-producing cell-stimulating factor activity. J Immunol. 1983 Jul;131(1):282–287. [PubMed] [Google Scholar]
  7. Inoue T., Carsten A. L., Cronkite E. P., Kelley J. E. Separation and concentration of murine hematopoietic stem cells (CFUS) using a combination of density gradient sedimentation and counterflow centrifugal elutriation. Exp Hematol. 1981 Jul;9(6):563–572. [PubMed] [Google Scholar]
  8. Iscove N. N., Roitsch C. A., Williams N., Guilbert L. J. Molecules stimulating early red cell, granulocyte, macrophage, and megakaryocyte precursors in culture: similarity in size, hydrophobicity, and charge. J Cell Physiol Suppl. 1982;1:65–78. doi: 10.1002/jcp.1041130412. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Koizumi S., Fine R. L., Curt G. A., Griffin J. D., Chabner B. A. Enrichment of myeloid progenitor cells from normal human bone marrow using an immune-rosette technique. Exp Hematol. 1985 Jul;13(6):560–565. [PubMed] [Google Scholar]
  11. Nakahata T., Gross A. J., Ogawa M. A stochastic model of self-renewal and commitment to differentiation of the primitive hemopoietic stem cells in culture. J Cell Physiol. 1982 Dec;113(3):455–458. doi: 10.1002/jcp.1041130314. [DOI] [PubMed] [Google Scholar]
  12. Nakahata T., Ogawa M. Clonal origin of murine hemopoietic colonies with apparent restriction to granuclocyte-macrophage-megakaryocyte (GMM) differentiation. J Cell Physiol. 1982 Jun;111(3):239–246. doi: 10.1002/jcp.1041110304. [DOI] [PubMed] [Google Scholar]
  13. Nakahata T., Ogawa M. Hemopoietic colony-forming cells in umbilical cord blood with extensive capability to generate mono- and multipotential hemopoietic progenitors. J Clin Invest. 1982 Dec;70(6):1324–1328. doi: 10.1172/JCI110734. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Nakahata T., Spicer S. S., Cantey J. R., Ogawa M. Clonal assay of mouse mast cell colonies in methylcellulose culture. Blood. 1982 Aug;60(2):352–361. [PubMed] [Google Scholar]
  16. Nicola N. A., Metcalf D., von Melchner H., Burgess A. W. Isolation of murine fetal hemopoietic progenitor cells and selective fractionation of various erythroid precursors. Blood. 1981 Aug;58(2):376–386. [PubMed] [Google Scholar]
  17. Prystowsky M. B., Otten G., Naujokas M. F., Vardiman J., Ihle J. N., Goldwasser E., Fitch F. W. Multiple hemopoietic lineages are found after stimulation of mouse bone marrow precursor cells with interleukin 3. Am J Pathol. 1984 Nov;117(2):171–179. [PMC free article] [PubMed] [Google Scholar]
  18. Quesenberry P. J., Ihle J. N., McGrath E. The effect of interleukin 3 and GM-CSA-2 on megakaryocyte and myeloid clonal colony formation. Blood. 1985 Jan;65(1):214–217. [PubMed] [Google Scholar]
  19. Rennick D. M., Lee F. D., Yokota T., Arai K. I., Cantor H., Nabel G. J. A cloned MCGF cDNA encodes a multilineage hematopoietic growth factor: multiple activities of interleukin 3. J Immunol. 1985 Feb;134(2):910–914. [PubMed] [Google Scholar]
  20. Suda J., Suda T., Ogawa M. Analysis of differentiation of mouse hemopoietic stem cells in culture by sequential replating of paired progenitors. Blood. 1984 Aug;64(2):393–399. [PubMed] [Google Scholar]
  21. Suda T., Suda J., Ogawa M. Disparate differentiation in mouse hemopoietic colonies derived from paired progenitors. Proc Natl Acad Sci U S A. 1984 Apr;81(8):2520–2524. doi: 10.1073/pnas.81.8.2520. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]
  23. 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]
  24. Suda T., Suda J., Ogawa M. Single-cell origin of mouse hemopoietic colonies expressing multiple lineages in variable combinations. Proc Natl Acad Sci U S A. 1983 Nov;80(21):6689–6693. doi: 10.1073/pnas.80.21.6689. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. TILL J. E., MCCULLOCH E. A., SIMINOVITCH L. A STOCHASTIC MODEL OF STEM CELL PROLIFERATION, BASED ON THE GROWTH OF SPLEEN COLONY-FORMING CELLS. Proc Natl Acad Sci U S A. 1964 Jan;51:29–36. doi: 10.1073/pnas.51.1.29. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Visser J. W., Bauman J. G., Mulder A. H., Eliason J. F., de Leeuw A. M. Isolation of murine pluripotent hemopoietic stem cells. J Exp Med. 1984 Jun 1;159(6):1576–1590. doi: 10.1084/jem.159.6.1576. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Williams N., Jackson H., Meyers P. Isolation of pluripotent hemopoietic stem cells and clonable precursor cells of erythrocytes, granulocytes, macrophages and megakaryocytes from mouse bone marrow. Exp Hematol. 1979 Nov;7(10):524–534. [PubMed] [Google Scholar]

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