Mouse epidermal keratinocytes in three-dimensional organotypic coculture with dermal fibroblasts form a stratified sheet resembling skin.
We describe an organotypic model of mouse skin consisting of a stratified sheet of epidermal keratinocytes and dermal fibroblasts within a contracted collagen gel. The model was designed to maintain the polarity of stratified keratinocytes and permit their long-term culture at an air-liquid interface. After air exposure, the thickness of the keratinocyte sheet transiently increased and then decreased to two cell layers at 2 weeks. The two-cell-layer structure is similar to that of the adult mouse epidermis. Cytokeratin 5 was localized in the lowest cell layer in the epithelial sheet, but cytokeratin 1 and loricrin were localized in the outer cell layers, resembling mouse skin. The expressions of interleukin 1alpha and 1beta in the keratinocytes and of keratinocyte growth factor 1 and 2 in the fibroblasts correlated with keratinocyte stratification. The mouse organotypic coculture is useful in studying epithelial cell-mesenchymal cell interactions in vitro.
Ikuta
Shuzo
S
Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan.
Sekino
Nobufumi
N
Hara
Takeshi
T
Saito
Yuriko
Y
Chida
Kazuhiro
K
D000818:Animals; D002454:Cell Differentiation; D049109:Cell Proliferation; D018920:Coculture Techniques; D016207:Cytokines / Q000235:genetics; D004247:DNA / Q000096:biosynthesis; D005347:Fibroblasts; D015603:Keratinocytes / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D008813:Mice, Inbred ICR; D012867:Skin / Q000166:cytology* / Q000378:metabolism; D023822:Tissue Engineering
D016207:Cytokines; D004247:DNA