Pancreatic Cancer and Breast Tumor Dissociation Protocol

At the end of treatment, pancreatic cancer tumor tissues were minced into small pieces with scissors and digested in DMEM containing 0.5 mg mL⁻¹ Collagenase IV (Yeasen), 0.5 mg mL⁻¹ Collagenase I (Yeasen), and 25 µg mL⁻¹ DNAse 1. (Aladdin). After 1 h of incubation at 37 °C, samples were ground and filtered through 70 µm cell strainers, then centrifuged at 1800 rpm for 5 min at 4 °C. Orthotopic 4T1 tumors were minced into small pieces with scissors and digested in DMEM containing 0.5 mg mL⁻¹ Collagenase IV (Yeasen) and 0.5 mg mL⁻¹ Collagenase I (Yeasen). The samples were incubated at 37 °C for 1 h 30 min, samples were ground and filtered through 70 µm cell strainers, then centrifuged at 1800 rpm for 5 min at 4 °C. The collected single‐cell suspensions were stained with fluorescence‐labeled antibodies. Flow cytometry using an Attune NxT device (Thermo Fisher Scientific) and analyzed by FlowJo software 10. Fluorescence‐conjugated antibodies used in flow cytometry were summarized in Table S1 (Supporting Information).