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In short, tissues were incubated with PBS containing EDTA at 37 C for 2 20 min, then digested twice for 35 min with a cocktail of collagenase D (1 mg/ml, Roche), dispase I (0.1 mg/ml, Roche), and DNase I (333 mg/ml, Sigma), smashed and filtered through a 40 mm gauze (BD Biosciences)