Fresh tissues were washed three times with Hank’s solution containing 1% FBS, cut into small pieces, collected in RPMI 1640 containing collagenase IV (1 mg/ml) and deoxyribonuclease I (10 mg/ml), and then mechanically dissociated by using the gentle MACS Dissociator (Miltenyi Biotec). Dissociated cells were further incubated for 0.5 to 1 hour at 37°C under continuous rotation. The cell suspensions were then filtered through a 70-μm cell strainer (BD Labware).

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