Briefly, HaCaT keratinocytes were seeded at a density of 10,000 cells per well in 200 l fresh culture medium (DMEM supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin solution) into 96-well flat-bottomed microtitre plates (Costar, Corning, USA) and incubated at 37 C in humidified 5% CO 2 using AutoFlow NU-5510 Direct Heat CO 2 Incubator (NuAire, USA) for 24 hours before treatment
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No other vial size comes close to that duration at the lower end of the dosing range
In our hands, minipig microsomes demonstrated a close representation of human in vivo metabolism with the successful hydrolysis of acetanilide, flutamide, lidocaine, prilocaine and propanil (Fig