Anti epcam magnetic beads

KPV^+/+ and KPV^−/− mice were treated with Ad-Cre as described above; after 6 weeks, mice were sacrificed and lung tumors were excised. The tissue was dissociated into a single-cell suspension in 0.2 mg/mL DNase and 2 mg/mL collagenase D and filtered through a 40 μm filter. Cells then underwent two rounds of selection. First, cells were treated with anti-CD45 magnetic beads (Miltenyi Biotec, 130-052-301) and were passed through a magnetic column. CD45-negative cells were then subjected to anti-EpCAM magnetic beads (Miltenyi Biotec, 130-105-958) and underwent positive selection. CD45-negative, EpCAM-positive cells were expanded in vitro and were used in experiments between passages 1 and 10. Cells derived from a human lung adenocarcinoma (A549) were obtained from the American Type Culture Collection (ATCC, Manassas, VA). All cells were maintained in Dulbecco’s modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum, 100 U/mL penicillin, 100 µg/mL streptomycin, and HEPES buffer. All cells were grown in a humidified incubator of 5% CO₂/95% air at 37 °C.