title: Human adult epidermal melanocytes cultured without chemical mitogens express the EGF receptor and respond to EGF.
abstract: We describe a novel chemical mitogen-free in vitro culture technique for obtaining pure melanocyte cultures using normal human adult epidermis as a source. The culture medium consists equal parts of the commercially available Keratinocyte Basal and AIM-V media (both from Gibco), as basal medium, which is supplemented with fetal bovine serum, bovine pituitary extract and recombinant human epidermal growth factor (EGF). Melanocytes harvested from human adult skin proliferate extensively and can be passaged serially up to 10-15 times using this medium. We have verified the identity of the cultured cells by tyrosinase mRNA expression and TRP-1 protein staining. Moreover, we showed that autologous human serum alone, without additional supplements is able to provide sufficient growth support for the cultured cells in the basal medium, making this culture technique suitable for autologous melanocyte transplantation. In this culture system normal human adult melanocytes expressed both EGF receptor (EGFR) mRNA and protein and EGF showed a dose dependent mitogenic effect on the cells. EGF itself had no significant influence on EGFR mRNA expression.
authors.affiliation: Faculty of Medicine, Department of Dermatology and Allergology, University of Szeged, 6720 Szeged, Korányi fasor 6, Hungary.
authors.forename: Gábor
authors.initials: G
authors.lastname: Szabad
authors.forename: Bernadett
authors.initials: B
authors.lastname: Kormos
authors.forename: Andor
authors.initials: A
authors.lastname: Pivarcsi
authors.forename: Márta
authors.initials: M
authors.lastname: Széll
authors.forename: Kornélia
authors.initials: K
authors.lastname: Kis
authors.forename: Anna
authors.initials: A
authors.lastname: Kenderessy Szabó
authors.forename: Attila
authors.initials: A
authors.lastname: Dobozy
authors.forename: Lajos
authors.initials: L
authors.lastname: Kemény
authors.forename: Zsuzsanna
authors.initials: Z
authors.lastname: Bata-Csörgo
mesh_terms: D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004815:Epidermal Growth Factor / Q000494:pharmacology*; D066246:ErbB Receptors / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008544:Melanocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D008934:Mitogens / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism
chemical_list: D008934:Mitogens; D012333:RNA, Messenger; D004815:Epidermal Growth Factor; D066246:ErbB Receptors