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Cells were seeded at a density of 5000 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
10.3390/ijms131215601 8 DoenhoffM
Semax has a very short serum half-life (minutes after systemic injection) because the base ACTH(4-10) sequence is rapidly degraded by enkephalinase
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