Preclinical studies frequently lack predictive value for human conditions. Human cell-based disease models that reflect patient heterogeneity may reduce the high failure rates of preclinical research. Herein, we investigated the impact of primary cell age and body region on skin homeostasis, epidermal differentiation, and drug uptake. Fibroblasts derived from the breast skin of female 20- to 30-year-olds or 60- to 70-year-olds and fibroblasts from juvenile foreskin (<10 years old) were compared in cell monolayers and in reconstructed human skin (RHS). RHS containing aged fibroblasts differed from its juvenile and adult counterparts, especially in terms of the dermal extracellular matrix composition and interleukin-6 levels. The site from which the fibroblasts were derived appeared to alter fibroblast-keratinocyte crosstalk by affecting, among other things, the levels of granulocyte-macrophage colony-stimulating factor. Consequently, the epidermal expression of filaggrin and e-cadherin was increased in RHS containing breast skin fibroblasts, as were lipid levels in the stratum corneum. In conclusion, the region of the body from which fibroblasts are derived appears to affect the epidermal differentiation of RHS, while the age of the fibroblast donors determines the expression of proteins involved in wound healing. Emulating patient heterogeneity in preclinical studies might improve the treatment of age-related skin conditions.

Charité Universitätsmedizin Berlin Charitéplatz 1 10117 Berlin Germany

Department of Molecular Biology University of Duisburg Essen Hufelandstr 55 45122 Essen Germany

Faculty of Pharmacy in Hradec Králové Charles University Akademika Heyrovského 1203 50005 Hradec Králové Czech Republic

Institute of Biomaterial Science Helmholtz Zentrum Geesthacht Kantstraße 55 14513 Teltow Germany

Institute of Nutritional Science Department of Nutritional Toxicology University of Potsdam Arthur Scheunert Allee 114 116 14558 Nuthetal Germany

Institute of Pharmacy Freie Universität Berlin Königin Luise Str 2 4 14195 Berlin Germany

IUF Leibniz Research Institute for Environmental Medicine Auf'm Hennekamp 50 40225 Düsseldorf Germany

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