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

Area of Science:

• Immunology
• Cancer Biology
• Molecular Biology

Background:

• Conventional type 1 dendritic cells (cDC1s) are crucial for anti-tumor immunity.
• cDC1s are often found in low numbers within the tumor microenvironment (TME).
• Understanding cDC1 function in the TME requires effective isolation and analysis methods.

Purpose of the Study:

• To establish a reliable protocol for identifying and isolating cDC1 subsets from murine implantable tumors.
• To enable subsequent transcriptomic profiling of isolated cDC1s.
• To facilitate deeper investigation into cDC1 roles in cancer immunity.

Main Methods:

• A flow sorting-based strategy was developed for cDC1 isolation.
• The protocol includes steps for tumor cell culture, dissociation, and extracellular staining.
• Procedures cover cell sorting, RNA isolation, mRNA library preparation, and sequencing.

Main Results:

• The protocol successfully isolates cDC1 subsets from murine tumors.
• This method allows for detailed transcriptomic analysis of tumor-infiltrating cDC1s.
• The study provides a reproducible workflow for cDC1 research.

Conclusions:

• The presented protocol is effective for isolating and analyzing cDC1s from the TME.
• This technique supports the study of cDC1s' innate sensing capabilities in cancer.
• The findings contribute to advancing cancer immunology research through improved cell isolation techniques.