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. Hypothesis/Objectives - 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. Animals - Four healthy beagle dogs from a research colony. Methods - Bulge cell-enriched keratinocytes showing keratin15 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 10-14days 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 keratin14 and desmoglein3 were detected in the basal and immediate suprabasilar layers of the epidermis, while keratin10 and desmoglein1 occurred in more superficial layers. Claudin1 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 and clinical importance - A bulge stem cell-enriched population from canine hair follicles formed interfollicular epidermis within 2weeks in vitro, and thus represents a promising model for regenerative therapy of canine skin.
| Original language | English |
|---|---|
| Journal | Veterinary Dermatology |
| Volume | 24 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - 2013 Feb |
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ASJC Scopus subject areas
- veterinary(all)
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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. / Kobayashi, Tetsuro; Enomoto, Kaoru; Wang, Yu Hsuan; Yoon, Ji Seon; Okamura, Ryoko; Ide, Kaori; Ohyama, Manabu; Nishiyama, Toshio; Iwasaki, Toshiroh; Nishifuji, Koji.
In: Veterinary Dermatology, Vol. 24, No. 1, 02.2013.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Epidermal structure created by canine hair follicle keratinocytes enriched with bulge cells in a three-dimensional skin equivalent model in vitro
T2 - Implications for regenerative therapy of canine epidermis
AU - Kobayashi, Tetsuro
AU - Enomoto, Kaoru
AU - Wang, Yu Hsuan
AU - Yoon, Ji Seon
AU - Okamura, Ryoko
AU - Ide, Kaori
AU - Ohyama, Manabu
AU - Nishiyama, Toshio
AU - Iwasaki, Toshiroh
AU - Nishifuji, Koji
PY - 2013/2
Y1 - 2013/2
N2 - 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. Hypothesis/Objectives - 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. Animals - Four healthy beagle dogs from a research colony. Methods - Bulge cell-enriched keratinocytes showing keratin15 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 10-14days 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 keratin14 and desmoglein3 were detected in the basal and immediate suprabasilar layers of the epidermis, while keratin10 and desmoglein1 occurred in more superficial layers. Claudin1 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 and clinical importance - A bulge stem cell-enriched population from canine hair follicles formed interfollicular epidermis within 2weeks in vitro, and thus represents a promising model for regenerative therapy of canine skin.
AB - 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. Hypothesis/Objectives - 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. Animals - Four healthy beagle dogs from a research colony. Methods - Bulge cell-enriched keratinocytes showing keratin15 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 10-14days 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 keratin14 and desmoglein3 were detected in the basal and immediate suprabasilar layers of the epidermis, while keratin10 and desmoglein1 occurred in more superficial layers. Claudin1 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 and clinical importance - A bulge stem cell-enriched population from canine hair follicles formed interfollicular epidermis within 2weeks in vitro, and thus represents a promising model for regenerative therapy of canine skin.
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UR - http://www.scopus.com/inward/citedby.url?scp=84872690020&partnerID=8YFLogxK
U2 - 10.1111/j.1365-3164.2012.01097.x
DO - 10.1111/j.1365-3164.2012.01097.x
M3 - Article
C2 - 23331683
AN - SCOPUS:84872690020
VL - 24
JO - Veterinary Dermatology
JF - Veterinary Dermatology
SN - 0959-4493
IS - 1
ER -