Division of Blood and Marrow Transplantation, Department of Medicine, University of California, San Diego (UCSD), La Jolla, CA, USA; Moores Cancer Center, University of California, San Diego (UCSD), La Jolla, CA, USA.
Division of Blood and Marrow Transplantation, Department of Medicine, University of California, San Diego (UCSD), La Jolla, CA, USA; Moores Cancer Center, University of California, San Diego (UCSD), La Jolla, CA, USA.
STAR Protoc. 2024 Sep 20;5(3):103195. doi: 10.1016/j.xpro.2024.103195. Epub 2024 Aug 1.
Conventional type 1 dendritic cells (cDC1s) are critical for innate sensing of cancer, yet they are scarce in the tumor microenvironment (TME). Here, we present a protocol to identify and isolate cDC1 subsets from murine implantable tumors for subsequent transcriptomic profiling using a flow sorting-based strategy. We describe steps for cell culture of mouse tumors, tumoral growth, dissociation and isolation of tumoral cells, extracellular staining, and cell sorting. We then detail procedures for RNA isolation, mRNA library preparation, and sequencing. For complete details on the use and execution of this protocol, please refer to Papadas et al..
传统的 1 型树突状细胞 (cDC1) 对于癌症的先天感知至关重要，但它们在肿瘤微环境 (TME) 中却很少。在这里，我们提出了一种使用基于流式分选的策略来鉴定和分离小鼠植入性肿瘤中的 cDC1 亚群并进行后续转录组分析的方案。我们描述了小鼠肿瘤细胞培养、肿瘤生长、肿瘤细胞解离和分离、细胞外染色和细胞分选的步骤。然后，我们详细介绍了 RNA 分离、mRNA 文库制备和测序的程序。有关该方案使用和执行的完整详细信息，请参阅 Papadas 等人的研究。
