title: Serum-free conditions for the long term growth and differentiation of neoplastic canine keratinocytes.
abstract: Long term cultures of canine keratinocytes have been established but culture conditions currently used require supplementation with fetal bovine serum (FBS). Unfortunately, FBS contains many non-defined components which may interfere with in vitro studies. This study describes the development of defined serum-free culture conditions for neoplastic canine keratinocytes grown submerged and at the air-liquid interface. Two commercially available serum-free media established for human epidermal cells failed to support canine keratinocyte growth. In contrast, a defined serum-free medium developed in our laboratory successfully supported proliferation of neoplastic canine keratinocytes for at least 40 passages. Cells showed a slower growth rate, but reached similar final densities and were morphologically identical to those cultured in FBS. Grown at the air-liquid interface, the cells reached the same degree of differentiation as in vivo stratified squamous epithelium and cultures grown in FBS. These results demonstrate that canine keratinocytes require different serum-free growth conditions than human cells. Neoplastic canine keratinocyte cultures, grown under serum-free culture conditions, provide an ideal in vitro system for comparative studies of keratinocyte biology and pathogenesis of various dermatoses.
authors.affiliation: Department of Pathology, College of Veterinary Medicine, Cornell University, Ithaca, NY, USA.
authors.forename: F M
authors.initials: FM
authors.lastname: Crameri
authors.forename: M
authors.initials: M
authors.lastname: Varvayanis
authors.forename: B R
authors.initials: BR
authors.lastname: Cromie
authors.forename: W L
authors.initials: WL
authors.lastname: Rekers
authors.forename: M M
authors.initials: MM
authors.lastname: Suter
mesh_terms: D000388:Air; D000818:Animals; D000943:Antigens, Differentiation / Q000032:analysis; D002294:Carcinoma, Squamous Cell / Q000473:pathology / Q000662:veterinary*; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D016895:Culture Media, Serum-Free / Q000494:pharmacology*; D004283:Dog Diseases / Q000473:pathology*; D004285:Dogs; D005434:Flow Cytometry; D006801:Humans; D015603:Keratinocytes / Q000187:drug effects / Q000473:pathology*; D037102:Lectins / Q000032:analysis; D009062:Mouth Neoplasms / Q000473:pathology / Q000662:veterinary*; D009363:Neoplasm Proteins / Q000032:analysis; D011975:Receptors, Mitogen / Q000032:analysis; D014407:Tumor Cells, Cultured
chemical_list: D000943:Antigens, Differentiation; D016895:Culture Media, Serum-Free; D037102:Lectins; D009363:Neoplasm Proteins; D011975:Receptors, Mitogen