Protocol

Basic Cell Culture Protocols

Volume 946 of the series Methods in Molecular Biology pp 85-102

Date:

The Development of T Lymphocytes in Fetal Thymus Organ Culture

  • Takeshi NittaAffiliated withDivision of Experimental Immunology, Institute for Genome Research, University of Tokushima
  • , Izumi OhigashiAffiliated withDivision of Experimental Immunology, Institute for Genome Research, University of Tokushima
  • , Yousuke TakahamaAffiliated withDivision of Experimental Immunology, Institute for Genome Research, The University of Tokushima Email author 

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Abstract

Fetal thymus organ culture (FTOC) is a unique and powerful culture system that allows intrathymic T-lymphocyte development in vitro. T-cell development in FTOC well represents fetal thymocyte development in vivo. Here we describe the basic method for FTOC as well as several related techniques, including reconstitution of thymus lobes with T-lymphoid progenitor cells, high-oxygen submersion culture, reaggregation thymus organ culture, retrovirus-mediated gene transfer to developing thymocytes in FTOC, and coculture of progenitor cells with OP9-DL1 cells.

Key words

T lymphocyte Fetal thymus Organ culture Retrovirus Flow cytometry