Retroviral Gene Transduction into T Cell Progenitors for Analysis of T Cell Development in the Thymus
Retroviral Gene Transduction into T Cell Progenitors for Analysis of T Cell Development in the Thymus
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DOI：
10.1007/978-1-0716-0266-9_16
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发表时间：
2020-01-01
期刊：
影响因子：
--
通讯作者：
Nitta, Takeshi
中科院分区：
文献类型：
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作者：
Muro, Ryunosuke;Takayanagi, Hiroshi;Nitta, Takeshi
胸腺是 T 细胞一生中发育的器官。使用小鼠作为模型动物，研究了胸腺内 T 细胞发育的分子机制。胎儿胸腺器官培养技术能够离体重建胎儿特异性T细胞发育，而骨髓嵌合技术允许体内重建成人胸腺中的T细胞发育。这些技术可以与 T 细胞祖细胞中的逆转录病毒基因转导相结合，以评估发育中 T 细胞中感兴趣的基因的功能。在这里，我们描述了逆转录病毒基因转导到胎儿或成人 T 细胞祖细胞和重建胸腺 T 细胞发育的基本方案，包括实验技巧，例如使用冷冻保存的胎儿肝脏或骨髓细胞作为 T 细胞祖细胞的来源。
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 Tcell development including experimental tips such as using cryopreserved fetal liver or bone marrow cells as sources of T cell progenitors.