He Zehua, Hu Ziyi, Wang Lei, Xiao Yibei, Cao Xu
Department of Pharmacology, School of Pharmacy, China Pharmaceutical University, Nanjing, China; Shanghai Frontiers Science Center for Drug Target Identification and Delivery, and the Engineering Research Center of Cell and Therapeutic Antibody of the Ministry of Education, National Key Laboratory of Innovative Immunotherapy, School of Pharmaceutical Sciences, Shanghai Jiao Tong University, Shanghai, China.
Shanghai Frontiers Science Center for Drug Target Identification and Delivery, and the Engineering Research Center of Cell and Therapeutic Antibody of the Ministry of Education, National Key Laboratory of Innovative Immunotherapy, School of Pharmaceutical Sciences, Shanghai Jiao Tong University, Shanghai, China.
STAR Protoc. 2025 Mar 21;6(1):103512. doi: 10.1016/j.xpro.2024.103512. Epub 2024 Dec 18.
Macrophage-mediated phagocytosis has emerged as a pivotal mechanism for eliminating tumor cells within the realm of cancer immunotherapy. Here, we present a protocol for identifying small molecules that enhance phagocytosis in mice using a co-culture system comprising primary macrophages, cancer cells, and a blockade of phagocytic checkpoints. We describe steps for expressing enhanced green fluorescent protein-luciferase (eGFP-Luc) and producing bone marrow-derived macrophages (BMDMs). We then detail procedures for optimizing co-culture conditions for high-throughput screen (HTS) and executing HTS chemical identification. For complete details on the use and execution of this protocol, please refer to Cao et al. .
巨噬细胞介导的吞噬作用已成为癌症免疫治疗领域中消除肿瘤细胞的关键机制。在此，我们展示了一种使用包含原代巨噬细胞、癌细胞和吞噬检查点阻断剂的共培养系统来鉴定增强小鼠吞噬作用的小分子的方案。我们描述了表达增强型绿色荧光蛋白 - 荧光素酶（eGFP-Luc）和产生骨髓来源巨噬细胞（BMDM）的步骤。然后，我们详细介绍了为高通量筛选（HTS）优化共培养条件以及进行HTS化学鉴定的程序。有关本方案使用和执行的完整详细信息，请参考Cao等人的研究。
