title: Epidermal structure created by canine hair follicle keratinocytes enriched with bulge cells in a three-dimensional skin equivalent model in vitro: implications for regenerative therapy of canine epidermis.
abstract: BACKGROUND
  Keratinocytes in the hair follicle bulge region have a high proliferative capacity, with characteristics of epithelial stem cells. This cell population might thus be an ideal source for generating the interfollicular epidermis in a canine skin equivalent.
  
  
  OBJECTIVE
  This study was designed to determine the ability of canine hair follicle bulge cell-enriched keratinocytes to construct canine living skin equivalents with interfollicular epidermis in vitro.
  
  
  METHODS
  Four healthy beagle dogs from a research colony.
  
  
  METHODS
  Bulge cell-enriched keratinocytes showing keratin 15 immunoreactivity were isolated from canine hair follicles and cultured on dermal equivalent containing canine fibroblasts. Skin equivalents were subjected to histological, immunohistochemical, western blot and RT-PCR analyses after 10-14 days of culture at the air-liquid interface.
  
  
  RESULTS
  The keratinocyte sheets showed an interfollicular epidermal structure comprising four to five living cell layers covered with a horny layer. Immunoreactivities for keratin 14 and desmoglein 3 were detected in the basal and immediate suprabasilar layers of the epidermis, while keratin 10 and desmoglein 1 occurred in more superficial layers. Claudin 1 immunoreactivity was seen in the suprabasalar layer of the constructed epidermis, and filaggrin monomers and loricrin were detected in the uppermost layer. Basal keratinocytes in the skin equivalent demonstrated immunoreactivity to antibodies against basement membrane zone molecules.
  
  
  CONCLUSIONS
  A bulge stem cell-enriched population from canine hair follicles formed interfollicular epidermis within 2 weeks in vitro, and thus represents a promising model for regenerative therapy of canine skin.
authors: Department of Dermatology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku, Tokyo, Japan.
authors: Tetsuro
authors: T
authors: Kobayashi
authors: Kaoru
authors: K
authors: Enomoto
authors: Yu Hsuan
authors: YH
authors: Wang
authors: Ji Seon
authors: JS
authors: Yoon
authors: Ryoko
authors: R
authors: Okamura
authors: Kaori
authors: K
authors: Ide
authors: Manabu
authors: M
authors: Ohyama
authors: Toshio
authors: T
authors: Nishiyama
authors: Toshiroh
authors: T
authors: Iwasaki
authors: Koji
authors: K
authors: Nishifuji
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000662:veterinary*; D004283:Dog Diseases / Q000628:therapy*; D004285:Dogs; D018859:Hair Follicle / Q000166:cytology*; D015603:Keratinocytes / Q000502:physiology*; D012871:Skin Diseases / Q000628:therapy / Q000662:veterinary*; D023822:Tissue Engineering / Q000379:methods / Q000662:veterinary*