常驻记忆 T 细胞 (TRM) 在区域免疫防御中发挥着至关重要的作用。尽管实验室啮齿动物已被广泛用于研究基本的 TRM 生物学，但较差的分离效率和低细胞存活率限制了以 TRM 为重点的高通量测定的实施。在这里，我们设计了一种支持 CD8 TRM 分化的小鼠阴道上皮类器官 (VEO)-CD8 T 细胞共培养系统。这些体外生成的 TRM 在表型和转录上与体内 TRM 相似。药理学和遗传学方法表明，转化生长因子 β (TGF-β) 信号在其分化中起着至关重要的作用。我们模型中的 VEO 容易受到病毒感染，而 CD8 T 细胞易于进行基因操作，这两者都将允许详细研究抗病毒 CD8 T 细胞生物学。总之，我们已经建立了一个强大的体外 TRM 分化系统，该系统是可扩展的，可以进行高通量测定，这将迅速增加我们对 TRM 的理解。版权所有 © 2024 作者。由爱思唯尔公司出版。保留所有权利。

Resident memory T cells (TRMs) play a vital role in regional immune defense. Although laboratory rodents have been extensively used to study fundamental TRM biology, poor isolation efficiency and low cell survival rates have limited the implementation of TRM-focused high-throughput assays. Here, we engineer a murine vaginal epithelial organoid (VEO)-CD8 T cell co-culture system that supports CD8 TRM differentiation. These in-vitro-generated TRMs are phenotypically and transcriptionally similar to in vivo TRMs. Pharmacological and genetic approaches showed that transforming growth factor β (TGF-β) signaling plays a crucial role in their differentiation. The VEOs in our model are susceptible to viral infections and the CD8 T cells are amenable to genetic manipulation, both of which will allow a detailed interrogation of antiviral CD8 T cell biology. Altogether we have established a robust in vitro TRM differentiation system that is scalable and can be subjected to high-throughput assays that will rapidly add to our understanding of TRMs.Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.