title: Effects of serially passaged fibroblasts on dermal and epidermal morphogenesis in human skin equivalents.
abstract: Serial passaging has a profound effect on primary cells. Since serially passaged cells show signs of cellular aging, serial passaging is used as an in vitro model of aging. To relate the effect of in vitro aging more to in vivo aging, we generated human skin equivalents (HSEs). We investigated if HSEs generated with late passage fibroblasts show characteristics of aged skin when compared with HSEs generated with early passage fibroblasts. Late passage fibroblasts had enlarged cell bodies and were more often positive for myofibroblast marker α-smooth muscle actin, senescence associated β-galactosidase and p16 compared with early passage fibroblasts. Skin equivalents generated with late passage fibroblasts had a thinner dermis, which could partly be explained by increased matrix metalloproteinase-1 secretion. In equivalents generated with late passage fibroblasts epidermal expression of keratin 6 was increased, and of keratin 10 slightly decreased. However, epidermal proliferation, epidermal thickness and basement membrane formation were not affected. In conclusion, compared with HSEs generated with early passage fibroblasts, HSEs generated with late passage fibroblasts showed changes in the dermis, but no or minimal changes in the basement membrane and the epidermis.
authors.affiliation: Department of Dermatology, Leiden University Medical Center, Room T-02-09, Postal Zone S2-P, P.O. Box 9600, 2300 RC, Leiden, The Netherlands. d.g.janson@lumc.nl
authors.forename: David
authors.initials: D
authors.lastname: Janson
authors.forename: Marion
authors.initials: M
authors.lastname: Rietveld
authors.forename: Rein
authors.initials: R
authors.lastname: Willemze
authors.forename: Abdoelwaheb
authors.initials: A
authors.lastname: El Ghalbzouri
mesh_terms: D000199:Actins / Q000378:metabolism; D000328:Adult; D002478:Cells, Cultured; D016922:Cellular Senescence / Q000502:physiology; D019941:Cyclin-Dependent Kinase Inhibitor p16; D020405:Dermis / Q000166:cytology / Q000254:growth & development*; D000078404:Epidermal Cells; D004817:Epidermis / Q000254:growth & development*; D005260:Female; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D053550:Keratin-10 / Q000378:metabolism; D053553:Keratin-6 / Q000378:metabolism; D015603:Keratinocytes / Q000166:cytology* / Q000378:metabolism; D020781:Matrix Metalloproteinase 1 / Q000378:metabolism; D008875:Middle Aged; D009024:Morphogenesis / Q000502:physiology*; D009363:Neoplasm Proteins / Q000378:metabolism; D012867:Skin / Q000166:cytology / Q000254:growth & development*; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: C541116:ACTA2 protein, human; D000199:Actins; C000614131:CDKN2A protein, human; D019941:Cyclin-Dependent Kinase Inhibitor p16; D053553:Keratin-6; D009363:Neoplasm Proteins; D053550:Keratin-10; D001616:beta-Galactosidase; D020781:Matrix Metalloproteinase 1