An optimized method for intensive screening of molecules that stimulate beta-defensin 2 or 3 (hBD2 or hBD3) expression in cultured normal human keratinocytes

摘要

Normal human skin controls the intrusion of microorganisms by the production of peptide antibiotics such as defensins.The aim of our study was to develop a culture model of normal human keratinocytes for optimal beta-defensin mRNA detection which allows the screening of molecules able to stimulate hBD2 and hBD3 without inducing pro-inflammatory cytokines.A keratinocyte culture model in 96-well plates,in high calcium medium (1.7 mM) allowed to analyze hBD2 and hBD3 mRNA expression in basal condition and after cell stimulation by products from diverse vegetal extracts.