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通过流式细胞术检测衰老肿瘤细胞刺激小鼠骨髓来源树突状细胞能力的方案。

Protocol for examining the capability of senescent tumor cells to stimulate murine bone-marrow-derived dendritic cells by flow cytometry.

机构信息

Ludwig Center for Metastasis Research and Department of Molecular Genetics and Cell Biology, The University of Chicago, Chicago, IL 60637, USA.

出版信息

STAR Protoc. 2023 Dec 15;4(4):102677. doi: 10.1016/j.xpro.2023.102677. Epub 2023 Oct 31.

Abstract

Therapy-induced senescence (TIS) may contribute to therapy resistance; however, evidence also suggests that senescent cells (SnCs) may promote anti-tumor immunity. Here, we present a protocol for examining the capability of TIS to stimulate type 1 conventional CD103 dendritic cells (DCs). We describe steps for isolating and differentiating CD103 DCs from murine bone marrow, inducing senescence in murine colon carcinoma cell line CT26, and coculturing DCs with SnCs. We then detail the flow cytometric analysis of DC maturation and activation. For complete details on the use and execution of this protocol, please refer to Liu et al. (2022) and Liu et al. (2023)..

摘要

治疗诱导的衰老(TIS)可能会导致治疗抵抗;然而,有证据表明衰老细胞(SnC)也可能促进抗肿瘤免疫。在这里,我们提出了一个方案,用于检查 TIS 刺激 1 型常规 CD103 树突状细胞(DC)的能力。我们描述了从鼠骨髓中分离和分化 CD103 DC 的步骤,诱导鼠结肠癌细胞系 CT26 的衰老,以及将 DC 与 SnC 共培养。然后,我们详细介绍了 DC 成熟和激活的流式细胞分析。有关该方案使用和执行的完整详细信息,请参阅 Liu 等人(2022 年)和 Liu 等人(2023 年)。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fe69/10751553/4b43b8dc6ec1/fx1.jpg

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