Isolation of intratumoral leukocytes of TLR-stimulated tumor-bearing mice.

Toll-like receptor (TLR) ligands hold promise for cancer immunotherapy. The isolation of intratumoral leukocytes of tumor-bearing mice is a useful technique for analyzing the immunological effects of TLR ligands on the tumor microenvironment. These isolated immune cells can be directly used for analysis (e.g., by flow cytometry) or cultured for functional in vitro studies. Here, we describe the isolation of intratumoral leukocytes by density gradient centrifugation. This technique can be used to isolate leukocytes from freshly dissected murine tumors.