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Differentiation of Mouse Thymocytes in Fetal Thymus Organ Culture

  • Protocol
T Cell Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 134))

Abstract

Most T cells develop in the thymus. Thymic development of T cells offers an excellent experimental system to pursue many issues of developmental biology, including

  1. 1.

    lineage decisions, e.g., CD4+ T cells vs CD8+ T cells, and TCR-αβ+ T cells vs TCR-γδ+ T cells,

  2. 2.

    clonal selection, e.g., positive selection of useful T-cell clones and negative selection of harmful T-cell clones, and

  3. 3.

    cell migration, e.g., entry into and emigration from the thymus.

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References

  1. Miller, J. F. A. P. (1961) Immunological function of the thymus. Lancet, 748–749.

    Google Scholar 

  2. Owen, J. J. T. and Ritter, M. A. (1969) Tissue interaction in the development of thymus lymphocytes. J. Exp. Med. 129, 431–442.

    Article  PubMed  CAS  Google Scholar 

  3. Owen, J. J. T. (1974) Ontogeny of the immune system. Prog. in Immunol. 2, 163–173.

    Google Scholar 

  4. Mandel, T. and Russel, P. J. (1971) Differentiation of foetal mouse thymus. ultrastructure of organ cultures and of subcapsular grafts. Immunology 21, 659–674.

    PubMed  CAS  Google Scholar 

  5. Mandel, T. E. and Kennedy, M. M. (1978) The differentiation of murine thymocytes in vivo and in vitro. Immunology 35, 317–331.

    PubMed  CAS  Google Scholar 

  6. Jenkinson, E. J., van Ewijk, W., and Owen, J. J. T. (1981) Major histocompatibility complex antigen expression on the epithelium of the developing thymus in normal and nude mice. J. Exp. Med. 153, 280–292.

    Article  PubMed  CAS  Google Scholar 

  7. Jenkinson, E. J., Franchi, L. L., Kingston, R., and Owen, J. J. T. (1982) Effect of deoxyguanosine on lymphopoiesis in the developing thymus rudiment in vitro: application in the production of chimeric thymus rudiments. Eur. J. Immunol. 12, 583–587.

    Article  PubMed  CAS  Google Scholar 

  8. Kingston, R., Jenkinson, E. J., and Owen, J. J. T. (1985) A single stem cell can recolonize an embryonic thymus, producing phenotypically distinct T-cell populations. Nature 317, 811–813.

    Article  PubMed  CAS  Google Scholar 

  9. Anderson, G., Jenkinson, E. J., Moore, N. C., and Owen, J. J. T. (1993) MHC class II-positive epithelium and mesenchyme cells are both required for T-cell development in the thymus. Nature 362, 70–73.

    Article  PubMed  CAS  Google Scholar 

  10. Kisielow, P., Leiserson, W., and von Boehmer, H. (1984) Differentiation of thymocytes in fetal organ culture: analysis of phenotypic changes accompanying the appearance of cytolytic and interleukin-2-producing cells. J. Immunol. 133, 1117–1123.

    PubMed  CAS  Google Scholar 

  11. Ceredig, R. (1988) Differentiation potential of 14-day fetal mouse thymocytes in organ culture. Analysis of CD4-CD8-defined single-positive and double-negative cells. J. Immunol. 141, 355–362.

    PubMed  CAS  Google Scholar 

  12. Husmann, L. A., Shimonkevitz, R. P., Crispe, I. N., and Bevan, M. J. (1988) Thymocyte subpopulations during early fetal development in the BALB/c mouse. J. Immunol. 141, 736–740.

    PubMed  CAS  Google Scholar 

  13. Takahama, Y., Hasegawa, T., Itohara, S., Ball, E. L., Sheard, M. A., and Hashimoto, Y. (1994) Entry of CD4−CD8− immature thymocytes into the CD4/CD8 developmental pathway is controlled by tyrosine kinase signals that can be provided through T cell receptor components. Int. Immunol. 6, 1505–1514.

    Article  PubMed  CAS  Google Scholar 

  14. Crompton, T., Gilmour, K. C., and Owen, M. J. (1996) The MAP kinase pathway controls differentiation from double-negative to double positive thymocyte. Cell 86, 243–251.

    Article  PubMed  CAS  Google Scholar 

  15. Heemskerk, M. H. M., Blom, B., Nolan, G., Stegmann, A. P. A., Bakker, A. Q., Weijer, K., Res, P. C. M., and Spits, H. (1997) Inhibition of T cell and promotion of natural killer cell development by the dominant negative helix loop helix factor Id3. J. Exp. Med. 186, 1597–1602.

    Article  PubMed  CAS  Google Scholar 

  16. Spain, L. M., Law, L. L., and Takahama, Y. (1998) Retroviral infection of T cell precursors in thymic organ culture, in Developmental Biology Protocols (Tuan, R. S. and Lo, C. W., eds.), Humana Press, Totowa, NJ, in press.

    Google Scholar 

  17. Sugawara, T., Di Bartolo, V., Miyazaki, T., Nakauchi, H., Acuto, O., and Takahama, Y. (1998) An improved retroviral transfer technique demonstrates inhibition of CD4−CD8− thymocyte development by kinase-inactive ZAP-70. J. Immunol. 161, 2888–2894.

    PubMed  CAS  Google Scholar 

  18. Ikuta, K., Kina, T., MacNeil, I., Uchida, N., Peault, B., Chien, Y. H., and Weissman, I. L. (1990) A developmental switch in thymic lymphocyte maturation potential occurs at the level of hematopoietic stem cells. Cell 62, 863–874.

    Article  PubMed  CAS  Google Scholar 

  19. Tsuda, S., Rieke, S., Hashimoto, Y., Nakauchi, H., and Takahama, Y. (1996) IL-7 supports D-J but not V-DJ rearrangement of T cell receptor β gene in fetal liver progenitor cells. J. Immunol. 156, 3233–3242.

    PubMed  CAS  Google Scholar 

  20. Sagara, S., Sugaya, K., Tokoro, Y., Tanaka, S., Takano, H., Kodama, H., Nakauchi, H., and Takahama, Y. (1997) B220 expression by T lymphoid progenitor cells in mouse fetal liver. J. Immunol. 158, 666–676.

    PubMed  CAS  Google Scholar 

  21. Theiler, K. (1989) The House Mouse. Springer-Verlag, New York, NY.

    Google Scholar 

  22. Kaufman, M. H. (1992) The Atlas of Mouse Development. Academic, San Diego, CA.

    Google Scholar 

  23. Butler, H. and Juurlink, B. H. (1987) An Atlas for Staging Mammalian and Chick Embryos. CRC, Boca Raton, FL.

    Google Scholar 

  24. Takahama, Y., Suzuki, H., Katz, K. S., Grusby, M. J., and Singer, A. (1994) Positive selection of CD4+ T cells by TCR ligation without aggregation even in the absence of MHC. Nature 371, 67–70.

    Article  PubMed  CAS  Google Scholar 

  25. Takahama, Y. and Nakauchi, H. (1996) Phorbol ester and calcium ionophore can replace TCR signals that induce positive selection of CD4 T cells. J. Immunol. 157, 1508–1513.

    PubMed  CAS  Google Scholar 

  26. Tokoro, Y., Tsuda, S., Tanaka, S., Nakauchi, H., and Takahama, Y. (1996) CD3-induced apoptosis of CD4+CD8+ thymocytes in the absence of clonotypic T-cell antigen receptor. Eur. J. Immunol. 26, 1012–1017.

    Article  PubMed  CAS  Google Scholar 

  27. Watanabe, Y. and Katsura, Y. (1993) Development of T cell receptor αβ-bearing T cells in the submersion organ culture of murine fetal thymus at high oxygen concentration. Eur. J. Immunol. 23, 200–205.

    Article  PubMed  CAS  Google Scholar 

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Author information

Authors and Affiliations

  1. University of Tsukuba, Japan

    Yousuke Takahama

Authors
  1. Yousuke Takahama
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Editor information

Editors and Affiliations

  1. Department of Microbiology and Immunology, East Carolina University School of Medicine, Greenville, NC

    Kelly P. Kearse

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© 2000 Humana Press Inc., Totowa, NJ

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Takahama, Y. (2000). Differentiation of Mouse Thymocytes in Fetal Thymus Organ Culture. In: Kearse, K.P. (eds) T Cell Protocols. Methods in Molecular Biology™, vol 134. Humana Press. https://doi.org/10.1385/1-59259-682-7:37

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  • DOI: https://doi.org/10.1385/1-59259-682-7:37

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-810-3

  • Online ISBN: 978-1-59259-682-9

  • eBook Packages: Springer Protocols

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