Serum-free conditions for the long term growth and differentiation of neoplastic canine keratinocytes.
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.
Crameri
F M
FM
Department of Pathology, College of Veterinary Medicine, Cornell University, Ithaca, NY, USA.
Varvayanis
M
M
Cromie
B R
BR
Rekers
W L
WL
Suter
M M
MM
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
D000943:Antigens, Differentiation; D016895:Culture Media, Serum-Free; D037102:Lectins; D009363:Neoplasm Proteins; D011975:Receptors, Mitogen