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These pieces were then incubated with digestion buffer (Dulbeccos modified Eagle medium with 2.5% FBS, 1% penicillin/streptomycin (Invitrogen, Carlsbad), 75 U/mL collagenase type IV (Gibco, Grand Island, NY), and 125 g/mL dispase type II (Gibco, Grand Island, NY)) at 37 C for 3060 min

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