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. 2022:2421:243-265.
doi: 10.1007/978-1-0716-1944-5_17.

Fetal Thymic Organ Culture (FTOC) Optimized for Gamma-Delta T Cell Studies

Johanna S Selvaratnam  1 , Tracy S H In  1 , Michele K Anderson  2
Affiliations

Fetal Thymic Organ Culture (FTOC) Optimized for Gamma-Delta T Cell Studies

Johanna S Selvaratnam et al. Methods Mol Biol. 2022.

Abstract

Fetal thymic organ culture (FTOC) provides a method for analyzing T cell development in a physiological context outside the animal. This technique enables studies of genetically altered mice that are embryonic or neonatal lethal, in addition to bypassing the complication of migration of successive waves of T cells out of the thymus. The hanging drop method involves depletion of thymocytes from host lobes using deoxyguanosine, followed by reconstitution with hematopoietic progenitors. This method has become standard for analysis of fetal liver precursors, bone marrow precursors, and early thymocytes. However, difficulties are encountered in the analysis of γδ T cell precursors using this method. We have developed a modification of FTOC in which partial depletion of hematopoietic precursors by shortened deoxyguanosine treatment, coupled with the use of TCRδ-deficient host lobes, enables engraftment and development of fetal γδTCR+ thymocytes. This method allows comparisons of development and functional differentiation of γδ T cell precursors between cells of different genotypes or treatments, in the context of a permissive thymic microenvironment.

Keywords: Development; FTOC; Flow cytometry; Gamma-delta T cells; Organ culture; Thymocytes; Thymus.

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References

    1. Auerbach R (1960) Morphogenetic interactions in the development of the mouse thymus gland. Dev Biol 2:271–284. https://doi.org/10.1016/0012-1606(60)90009-9 - DOI - PubMed
    1. Jenkinson EJ, Franchi LL, Kingston R, Owen JJ (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(7):583–587. https://doi.org/10.1002/eji.1830120710 - DOI - PubMed
    1. Anderson MK, Weiss AH, Hernandez-Hoyos G, Dionne CJ, Rothenberg EV (2002) Constitutive expression of PU.1 in fetal hematopoietic progenitors blocks T cell development at the pro-T cell stage. Immunity 16(2):285–296. https://doi.org/10.1016/s1074-7613(02)00277-7 - DOI - PubMed
    1. Anderson G, Jenkinson EJ, Moore NC, Owen JJ (1993) MHC class II-positive epithelium and mesenchyme cells are both required for T-cell development in the thymus. Nature 362(6415):70–73. https://doi.org/10.1038/362070a0 - DOI - PubMed
    1. Jenkinson EJ, Van Ewijk W, Owen JJ (1981) Major histocompatibility complex antigen expression on the epithelium of the developing thymus in normal and nude mice. J Exp Med 153(2):280–292. https://doi.org/10.1084/jem.153.2.280 - DOI - PubMed

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