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. 1991 May 1;173(5):1257–1266. doi: 10.1084/jem.173.5.1257

Different stromal cell lines support lineage-selective differentiation of the multipotential bone marrow stem cell clone LyD9

PMCID: PMC2118856  PMID: 1708811

Abstract

An interleukin 3-dependent multipotential stem cell clone, LyD9, has been shown to generate mature B lymphocytes, macrophages, and neutrophils by coculture with primary bone marrow stromal cells. We report here that coculture with the cloned stromal cell lines PA6 and ST2 can support differentiation of LyD9 cells predominantly into granulocyte/macrophage colony-stimulating factor (GM-CSF)- and granulocyte (G)-CSF-responsive cells, respectively. However, these stromal cell lines were unable to support lymphopoiesis of LyD9 cells. The GM-CSF-dependent line, L-GM, which was derived from LyD9 cells cocultured with PA6 stromal cells, could differentiate into macrophages and granulocytes in the presence of GM-CSF. The L-GM line can further differentiate predominantly into neutrophils by coculture with ST2 stromal cells. The G-CSF-dependent line, L-G, which was derived from LyD9 cells cocultured with ST2 stromal cells, differentiated into neutrophils in response to G-CSF. Although the stromal cell-supported differentiation of LyD9 cells required the direct contact between LyD9 and stromal cells, a small fraction of LyD9 cells that were pretreated with 5-azacytidine could differentiate into neutrophils and macrophages without direct contact with stromal cells. These results indicate that different stromal cell lines support lineage-selective differentiation of the LyD9 stem cell and that 5-azacytidine treatment can bypass the requirement of direct contact with stromal cells, albeit with a lower frequency.

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Selected References

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  1. Austyn J. M., Gordon S. F4/80, a monoclonal antibody directed specifically against the mouse macrophage. Eur J Immunol. 1981 Oct;11(10):805–815. doi: 10.1002/eji.1830111013. [DOI] [PubMed] [Google Scholar]
  2. Bentley S. A. Close range cell:cell interaction required for stem cell maintenance in continuous bone marrow culture. Exp Hematol. 1981 Mar;9(3):308–312. [PubMed] [Google Scholar]
  3. Castellazzi M., Vielh P., Longacre S. Azacytidine-induced reactivation of adenosine deaminase in a murine cytotoxic T cell line. Eur J Immunol. 1986 Sep;16(9):1081–1086. doi: 10.1002/eji.1830160909. [DOI] [PubMed] [Google Scholar]
  4. Coffman R. L., Weissman I. L. B220: a B cell-specific member of th T200 glycoprotein family. Nature. 1981 Feb 19;289(5799):681–683. doi: 10.1038/289681a0. [DOI] [PubMed] [Google Scholar]
  5. Constantinides P. G., Jones P. A., Gevers W. Functional striated muscle cells from non-myoblast precursors following 5-azacytidine treatment. Nature. 1977 May 26;267(5609):364–366. doi: 10.1038/267364a0. [DOI] [PubMed] [Google Scholar]
  6. Darmon M., Nicolas J. F., Lamblin D. 5-Azacytidine is able to induce the conversion of teratocarcinoma-derived mesenchymal cells into epithelia cells. EMBO J. 1984 May;3(5):961–967. doi: 10.1002/j.1460-2075.1984.tb01914.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dexter T. M., Allen T. D., Lajtha L. G. Conditions controlling the proliferation of haemopoietic stem cells in vitro. J Cell Physiol. 1977 Jun;91(3):335–344. doi: 10.1002/jcp.1040910303. [DOI] [PubMed] [Google Scholar]
  8. Dexter T. M., Garland J., Scott D., Scolnick E., Metcalf D. Growth of factor-dependent hemopoietic precursor cell lines. J Exp Med. 1980 Oct 1;152(4):1036–1047. doi: 10.1084/jem.152.4.1036. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dexter T. M., Spooncer E. Growth and differentiation in the hemopoietic system. Annu Rev Cell Biol. 1987;3:423–441. doi: 10.1146/annurev.cb.03.110187.002231. [DOI] [PubMed] [Google Scholar]
  10. Hara K., Suda T., Suda J., Eguchi M., Ihle J. N., Nagata S., Miura Y., Saito M. Bipotential murine hemopoietic cell line (NFS-60) that is responsive to IL-3, GM-CSF, G-CSF, and erythropoietin. Exp Hematol. 1988 May;16(4):256–261. [PubMed] [Google Scholar]
  11. Heyworth C. M., Dexter T. M., Kan O., Whetton A. D. The role of hemopoietic growth factors in self-renewal and differentiation of IL-3-dependent multipotential stem cells. Growth Factors. 1990;2(2-3):197–211. doi: 10.3109/08977199009071506. [DOI] [PubMed] [Google Scholar]
  12. Ihle J. N., Rebar L., Keller J., Lee J. C., Hapel A. J. Interleukin 3: possible roles in the regulation of lymphocyte differentiation and growth. Immunol Rev. 1982;63:5–32. doi: 10.1111/j.1600-065x.1982.tb00409.x. [DOI] [PubMed] [Google Scholar]
  13. Kajigaya S., Suda T., Suda J., Saito M., Miura Y., Iizuka M., Kobayashi S., Minato N., Sudo T. A recombinant murine granulocyte/macrophage (GM) colony-stimulating factor derived from an inducer T cell line (IH5.5). Functional restriction to GM progenitor cells. J Exp Med. 1986 Oct 1;164(4):1102–1113. doi: 10.1084/jem.164.4.1102. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Karasuyama H., Melchers F. Establishment of mouse cell lines which constitutively secrete large quantities of interleukin 2, 3, 4 or 5, using modified cDNA expression vectors. Eur J Immunol. 1988 Jan;18(1):97–104. doi: 10.1002/eji.1830180115. [DOI] [PubMed] [Google Scholar]
  15. Kinashi T., Inaba K., Tsubata T., Tashiro K., Palacios R., Honjo T. Differentiation of an interleukin 3-dependent precursor B-cell clone into immunoglobulin-producing cells in vitro. Proc Natl Acad Sci U S A. 1988 Jun;85(12):4473–4477. doi: 10.1073/pnas.85.12.4473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kinashi T., Lee K. H., Ogawa M., Tohyama K., Tashiro K., Fukunaga R., Nagata S., Honjo T. Premature expression of the macrophage colony-stimulating factor receptor on a multipotential stem cell line does not alter differentiation lineages controlled by stromal cells used for coculture. J Exp Med. 1991 May 1;173(5):1267–1279. doi: 10.1084/jem.173.5.1267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kinashi T., Tashiro K., Inaba K., Takeda T., Palacios R., Honjo T. An interleukin-4-dependent precursor clone is an intermediate of the differentiation pathway from an interleukin-3-dependent precursor clone into myeloid cells as well as B lymphocytes. Int Immunol. 1989;1(1):11–19. doi: 10.1093/intimm/1.1.11. [DOI] [PubMed] [Google Scholar]
  18. Kincade P. W., Lee G., Pietrangeli C. E., Hayashi S., Gimble J. M. Cells and molecules that regulate B lymphopoiesis in bone marrow. Annu Rev Immunol. 1989;7:111–143. doi: 10.1146/annurev.iy.07.040189.000551. [DOI] [PubMed] [Google Scholar]
  19. Kodama H. A., Amagai Y., Koyama H., Kasai S. Hormonal responsiveness of a preadipose cell line derived from newborn mouse calvaria. J Cell Physiol. 1982 Jul;112(1):83–88. doi: 10.1002/jcp.1041120113. [DOI] [PubMed] [Google Scholar]
  20. Nagata S., Tsuchiya M., Asano S., Kaziro Y., Yamazaki T., Yamamoto O., Hirata Y., Kubota N., Oheda M., Nomura H. Molecular cloning and expression of cDNA for human granulocyte colony-stimulating factor. 1986 Jan 30-Feb 5Nature. 319(6052):415–418. doi: 10.1038/319415a0. [DOI] [PubMed] [Google Scholar]
  21. Nicola N. A. Hemopoietic cell growth factors and their receptors. Annu Rev Biochem. 1989;58:45–77. doi: 10.1146/annurev.bi.58.070189.000401. [DOI] [PubMed] [Google Scholar]
  22. Nishikawa S., Ogawa M., Nishikawa S., Kunisada T., Kodama H. B lymphopoiesis on stromal cell clone: stromal cell clones acting on different stages of B cell differentiation. Eur J Immunol. 1988 Nov;18(11):1767–1771. doi: 10.1002/eji.1830181117. [DOI] [PubMed] [Google Scholar]
  23. Noma Y., Sideras P., Naito T., Bergstedt-Lindquist S., Azuma C., Severinson E., Tanabe T., Kinashi T., Matsuda F., Yaoita Y. Cloning of cDNA encoding the murine IgG1 induction factor by a novel strategy using SP6 promoter. Nature. 1986 Feb 20;319(6055):640–646. doi: 10.1038/319640a0. [DOI] [PubMed] [Google Scholar]
  24. Ogawa M., Nishikawa S., Ikuta K., Yamamura F., Naito M., Takahashi K., Nishikawa S. B cell ontogeny in murine embryo studied by a culture system with the monolayer of a stromal cell clone, ST2: B cell progenitor develops first in the embryonal body rather than in the yolk sac. EMBO J. 1988 May;7(5):1337–1343. doi: 10.1002/j.1460-2075.1988.tb02949.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Palacios R., Steinmetz M. Il-3-dependent mouse clones that express B-220 surface antigen, contain Ig genes in germ-line configuration, and generate B lymphocytes in vivo. Cell. 1985 Jul;41(3):727–734. doi: 10.1016/s0092-8674(85)80053-2. [DOI] [PubMed] [Google Scholar]
  26. Springer T., Galfré G., Secher D. S., Milstein C. Mac-1: a macrophage differentiation antigen identified by monoclonal antibody. Eur J Immunol. 1979 Apr;9(4):301–306. doi: 10.1002/eji.1830090410. [DOI] [PubMed] [Google Scholar]
  27. Takeda S., Gillis S., Palacios R. In vitro effects of recombinant interleukin 7 on growth and differentiation of bone marrow pro-B- and pro-T-lymphocyte clones and fetal thymocyte clones. Proc Natl Acad Sci U S A. 1989 Mar;86(5):1634–1638. doi: 10.1073/pnas.86.5.1634. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Tohyama K., Lee K. H., Tashiro K., Kinashi T., Honjo T. Establishment of an interleukin-5-dependent subclone from an interleukin-3-dependent murine hemopoietic progenitor cell line, LyD9, and its malignant transformation by autocrine secretion of interleukin-5. EMBO J. 1990 Jun;9(6):1823–1830. doi: 10.1002/j.1460-2075.1990.tb08307.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Unkeless J. C. Characterization of a monoclonal antibody directed against mouse macrophage and lymphocyte Fc receptors. J Exp Med. 1979 Sep 19;150(3):580–596. doi: 10.1084/jem.150.3.580. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Valtieri M., Tweardy D. J., Caracciolo D., Johnson K., Mavilio F., Altmann S., Santoli D., Rovera G. Cytokine-dependent granulocytic differentiation. Regulation of proliferative and differentiative responses in a murine progenitor cell line. J Immunol. 1987 Jun 1;138(11):3829–3835. [PubMed] [Google Scholar]
  31. Whitlock C. A., Witte O. N. Long-term culture of B lymphocytes and their precursors from murine bone marrow. Proc Natl Acad Sci U S A. 1982 Jun;79(11):3608–3612. doi: 10.1073/pnas.79.11.3608. [DOI] [PMC free article] [PubMed] [Google Scholar]

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