Bactericidal activity of flucloxacillin against Staphylococcus aureus in primary keratinocyte cultures of lesional and unaffected skin of patients suffering from atopic dermatitis.

More than 90% of patients with atopic dermatitis (AD) are colonized by Staphylococcus aureus. Due to prevalent multi-resistant strains, it is highly difficult to eliminate S. aureus contamination of primary cell cultures of human AD skin by means of the antibiotics commonly used in cell culture. Therefore, an anti-staphylococcal treatment was investigated by which sterile proliferating keratinocyte cultures are attained from lesional and unaffected skin of AD patients by applying flucloxacillin, which in general is systemically used in vivo. The treatment with 1% flucloxacillin for 20 min included both colonized skin samples and contaminated primary cell cultures. In the case of persistent contamination, this step was repeated as often as required until complete decontamination without any cytotoxic indications was achieved. Antibacterial treatment with flucloxacillin permitted the cultivation of sterile, vital, and proliferating primary cultures of human skin keratinocytes from lesional and unaffected skin of AD patients with S. aureus colonization. The method increased the success rate of isolation of AD keratinocytes from 30 to 90%, representing on average 3 vs. 9 successfully isolated, sterile and proliferating cultures out of 10 contaminated skin biopsies, which is comparable to normal human keratinocyte isolation rates.