Culturing Primary Human Keratinocytes

This study (project identification: No. 465) was approved on 23 February 2016 by Istituto Dermopatico dell’Immacolata review board, and performed according to the Declaration of Helsinki Guidelines. Primary cultures of normal human keratinocytes were obtained from healthy donors undergoing plastic surgery (n = 4, two females and two males, age 25–40) as previously described [40 (link)]. Keratinocytes were grown in serum-free Keratinocyte Growth Medium (Lonza, Walkersville, MD, USA), formed of Keratinocyte Basal Medium added of EGF, hydrocortisone, bovine insulin, bovine pituitary extract, and gentamycin sulfate [15 (link)]. In the 24 h before and during the experiments, keratinocyte cultures were kept in Keratinocyte Basal Medium (Lonza).