Protocol to identify and isolate rare murine tumor-resident dendritic cell populations for low-input transcriptomic profiling.

STAR Protoc

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. Electronic address:

Published: September 2024

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Article Abstract

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..

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC11345558PMC
http://dx.doi.org/10.1016/j.xpro.2024.103195DOI Listing

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