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.
