Retroviral Gene Transduction into T Cell Progenitors for Analysis of T Cell Development in the Thymus
The thymus is an organ where T cells develop throughout life. Using mice as a model animal, molecular mechanisms of intrathymic T cell development have been studied. Fetal thymus organ culture technique enables ex vivo reconstitution of fetal-specific T cell development, while bone marrow chimera technique allows in vivo reconstitution of T cell development in adult thymus. These techniques can be combined with retroviral gene transduction into the T cell progenitors to evaluate the function of genes of interest in developing T cells. Here, we describe the basic protocols for retrovirus gene transduction into fetal or adult T cell progenitors and reconstitution of thymic T cell development including experimental tips such as using cryopreserved fetal liver or bone marrow cells as sources of T cell progenitors.
Key wordsThymus T cells Retrovirus Fetal thymus organ culture Bone marrow chimera
We thank S. Nitta and M. Tsutsumi for the technical assistance. This study was supported by Grants-in-Aid for Research from the Japan Society for the Promotion of Science (JSPS) (KAKENHI 15H05703, 16H05202, 16K19102, and 17H05788), the Kanehara-Ichiro Foundation (grants 29–23), and the Kanae Foundation for the promotion of medical science (grant 47th).
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