Sarcomas were grown in mice until approximately 1 × 1 × 1 cm in size. Tumors were resected and enzymatically digested. Where applicable, tumor cells were FACS-sorted based on the expression of the fluorescent marker dsRED.
Human sarcoma samples were minced and then enzymatically and mechanically digested using the 37C_h_TDK_2 protocol on the Miltenyi gentleMACS with Tumor Dissociation Kit for Human cells (Miltenyi Biotec, Auburn, CA). Cell suspensions were washed in DPBS with 0.04% BSA and filtered through 70 μm strainers (Bioland Scientific LLC, Paramount, CA). Red blood cells were lysed with ACK buffer prior to staining with the following surface antibodies: anti-CD45 Pacific Blue, anti-CD11b APC, anti-CD163 PE, anti-CD3 FITC, anti-CD4 PE, anti-CD8 APC, anti-CD56 APC Cy7, anti-CD19 FITC (all purchased from Biolegend).
Human sarcoma samples were minced and then enzymatically and mechanically digested using the 37C_h_TDK_2 protocol on the Miltenyi gentleMACS with Tumor Dissociation Kit for Human cells (Miltenyi Biotec, Auburn, CA). Cell suspensions were washed in DPBS with 0.04% BSA and filtered through 70 μm strainers (Bioland Scientific LLC, Paramount, CA). Red blood cells were lysed with ACK buffer prior to staining with the following surface antibodies: anti-CD45 Pacific Blue, anti-CD11b APC, anti-CD163 PE, anti-CD3 FITC, anti-CD4 PE, anti-CD8 APC, anti-CD56 APC Cy7, anti-CD19 FITC (all purchased from Biolegend).