title: Compound screening and transcriptional profiling in human primary keratinocytes: a brief guideline.
abstract: Cultured human primary keratinocytes constitute suitable targets for in-depth evaluation of the proliferative or differentiative potential of compounds. There is, however, a double-edged and intrinsically inseparable transition from biological activity to cytotoxicity for any agent under investigation. For that reason, we here first of all present an established protocol for the isolation, cultivation, and analysis of primary foreskin-derived keratinocytes. Taking calcitriol as example, we then reveal how a straightforward photometric cell culture assay can be exploited to assess overall cell viability in response to increasing compound doses. With predetermined cellular cytotoxicity at hand, physiologically meaningful (sub-toxic) compound concentrations for subsequent stimulation of cells can be readily selected, and, in doing so, differentially expressed genes with biological significance can be reliably identified.
authors.affiliation: Division of Molecular Dermatology, Department of Dermatology, Paracelsus Private Medical University Salzburg, Salzburg, Austria.
authors.forename: Raphaela
authors.initials: R
authors.lastname: Rid
authors.forename: Harald
authors.initials: H
authors.lastname: Hundsberger
authors.forename: Kamil
authors.initials: K
authors.lastname: Onder
mesh_terms: D002117:Calcitriol / Q000494:pharmacology / Q000633:toxicity; D018929:Cell Culture Techniques; D049109:Cell Proliferation / Q000187:drug effects; D002469:Cell Separation; D002470:Cell Survival / Q000187:drug effects; D015925:Cryopreservation; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D012313:RNA; D002117:Calcitriol