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Flow Cytometry Analysis of Thymic Epithelial Cells and Their Subpopulations

  • Izumi Ohigashi
  • Yousuke TakahamaEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1323)

Abstract

The parenchyma of the thymus is compartmentalized into the cortex and the medulla, which are constructed by cortical thymic epithelial cells (cortical TECs, cTECs) and medullary thymic epithelial cells (mTECs), respectively. cTECs and mTECs essentially and differentially regulate the development and repertoire selection of T cells. Consequently, the biology of T cell development and selection includes the study of TECs in addition to the study of developing T cells and other hematopoietic cells including dendritic cells. In this chapter, we describe the methods for flow cytometric analysis and sorting of TECs and their subpopulations, including cTECs and mTECs.

Key words

Thymic epithelial cell mTEC cTEC Aire β5t Flow cytometry Cell sorting 

References

  1. 1.
    Petrie HT, Zúñiga-Pflücker JC (2007) Zoned out: functional mapping of stromal signaling microenvironments in the thymus. Annu Rev Immunol 25:649–679CrossRefPubMedGoogle Scholar
  2. 2.
    Klein L, Hinterberger M, Wirnsberger G, Kyewski B (2009) Antigen presentation in the thymus for positive selection and central tolerance induction. Nat Rev Immunol 9:833–844CrossRefPubMedGoogle Scholar
  3. 3.
    Anderson G, Takahama Y (2012) Thymic epithelial cells: working class heroes for T cell development and repertoire selection. Trends Immunol 33:256–263CrossRefPubMedGoogle Scholar
  4. 4.
    Alves NL, Goff OR, Huntington ND (2009) Characterization of the thymic IL-7 niche in vivo. Proc Natl Acad Sci U S A 106:1512–1517PubMedCentralCrossRefPubMedGoogle Scholar
  5. 5.
    Hara T, Shitara S, Imai K et al (2012) Identification of IL-7 producing cells in primary and secondary lymphoid organ using IL7-GFP knock-in mice. J Immunol 189:1577–1584CrossRefPubMedGoogle Scholar
  6. 6.
    Hozumi K, Mailhos C, Negishi N et al (2008) Delta-like 4 is indispensable in thymic environment specific for T cell development. J Exp Med 205:2507–2513PubMedCentralCrossRefPubMedGoogle Scholar
  7. 7.
    Koch U, Fiorini E, Benedito R et al (2008) Delta-like 4 is the essential, nonredundant ligand for Notch1 during thymic T cell lineage commitment. J Exp Med 205:2515–2523PubMedCentralCrossRefPubMedGoogle Scholar
  8. 8.
    Murata S, Sasaki K, Kishimoto T et al (2007) Regulation of CD8+ T cell development by thymus-specific proteasomes. Science 316:1349–1353CrossRefPubMedGoogle Scholar
  9. 9.
    Nitta T, Murata S, Sasaki K et al (2010) Thymoproteasome shapes immunocompetent repertoire of CD8+ T cells. Immunity 32:29–40CrossRefPubMedGoogle Scholar
  10. 10.
    Nagamine K, Peterson P, Scott HS et al (1997) Positional cloning of the APECED gene. Nat Genet 17:393–398CrossRefPubMedGoogle Scholar
  11. 11.
    Anderson MS, Venanzi ES, Klein L et al (2002) Projection of an immunological self shadow within the thymus by the Aire protein. Science 298:1395–1401CrossRefPubMedGoogle Scholar
  12. 12.
    Ueno T, Saito F, Gray DH et al (2004) CCR7 signals are essential for cortex-medulla migration of developing thymocytes. J Exp Med 200:493–505PubMedCentralCrossRefPubMedGoogle Scholar
  13. 13.
    Liu C, Saito F, Liu Z et al (2006) Coordination between CCR7- and CCR9-mediated chemokine signals in prevascular fetal thymus colonization. Blood 108:2531–2539CrossRefPubMedGoogle Scholar
  14. 14.
    Laufer TM, DeKoning J, Markowitz JS et al (1996) Unopposed positive selection and autoreactivity in mice expressing class II MHC only on thymic cortex. Nature 383:81–85CrossRefPubMedGoogle Scholar
  15. 15.
    Gray DH, Chidgey AP, Boyd RL (2002) Analysis of thymic stroma cell populations using flow cytometry. J Immunol Methods 260:15–28CrossRefPubMedGoogle Scholar
  16. 16.
    Williams KM, Mella H, Lucas PJ et al (2009) Single cell analysis of complex thymus stroma cell populations: rapid thymic epithelial preparation characterizes radiation injury. Clin Transl Sci 2:279–285PubMedCentralCrossRefPubMedGoogle Scholar
  17. 17.
    Nakagawa Y, Ohigashi I, Nitta T et al (2012) Thymic nurse cells provide microenvironment for secondary TCRα rearrangement in cortical thymocytes. Proc Natl Acad Sci U S A 109:20572–20577PubMedCentralCrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Division of Experimental Immunology, Institute for Genome ResearchUniversity of TokushimaTokushimaJapan

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