TY - JOUR
T1 - Reconstitution of human epidermis in vitro is accompanied by transient activation of basal keratinocyte spreading
AU - Grinnell, Frederick
AU - Toda, Ken Ichi
AU - Lamke-Seymour, Cheryl
N1 - Funding Information:
The studies were supported by NIH Grants GM31321 (to F.G.) and GM34513 (to Dr. Charles Baxter). We thank Drs. George Bloom, William Snell, and Woodring Wright for their helpful commentsr egardingt his manuscript.
PY - 1987/10
Y1 - 1987/10
N2 - Frozen human cadaver skin obtained from the skin bank was thawed and incubated in serum-free medium for 1-2 days, after which the original epidermis could be removed mechanically. Transmission electron microscopic observations showed that the dermal matrix remaining behind contained intact bundles of collagen fibrils but no live cells and that a continuous lamina densa persisted in the basement membrane region. Indirect immunofluorescence analyses demonstrated linear staining of the basement membrane region by antibodies against laminin and type IV collagen and discontinuous staining with antibodies against fibronectin. Scanning electron microscopic observations revealed a normal topographical arrangement of dermal matrix papilla and interspersed crypts on the surface of the matrix. Epidermal cells placed on the dermal matrix attached in 1-2 h and spread by 24 h. After 1 week of culture the epidermis was reconstituted, at which time approximately 30% of the epidermal cells were basal keratinocytes and the remainder were more differentiated keratinocytes. A high degree of differentiation of the reconstituted epidermis was shown by the formation of hemidesmosomes along the basement membrane, the formation of desmosomes characterized by intercellular dense lines, and the presence of a cell layer containing keratohyalin granules. At various times during epidermal reconstitution, cells were harvested and tested in short-term assays for adhesion to fibronectin substrata. During the first several days there was a transient activation of basal keratinocyte spreading analogous to the modulation of keratinocyte spreading that we have observed during epidermal reconstitution in vivo.
AB - Frozen human cadaver skin obtained from the skin bank was thawed and incubated in serum-free medium for 1-2 days, after which the original epidermis could be removed mechanically. Transmission electron microscopic observations showed that the dermal matrix remaining behind contained intact bundles of collagen fibrils but no live cells and that a continuous lamina densa persisted in the basement membrane region. Indirect immunofluorescence analyses demonstrated linear staining of the basement membrane region by antibodies against laminin and type IV collagen and discontinuous staining with antibodies against fibronectin. Scanning electron microscopic observations revealed a normal topographical arrangement of dermal matrix papilla and interspersed crypts on the surface of the matrix. Epidermal cells placed on the dermal matrix attached in 1-2 h and spread by 24 h. After 1 week of culture the epidermis was reconstituted, at which time approximately 30% of the epidermal cells were basal keratinocytes and the remainder were more differentiated keratinocytes. A high degree of differentiation of the reconstituted epidermis was shown by the formation of hemidesmosomes along the basement membrane, the formation of desmosomes characterized by intercellular dense lines, and the presence of a cell layer containing keratohyalin granules. At various times during epidermal reconstitution, cells were harvested and tested in short-term assays for adhesion to fibronectin substrata. During the first several days there was a transient activation of basal keratinocyte spreading analogous to the modulation of keratinocyte spreading that we have observed during epidermal reconstitution in vivo.
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U2 - 10.1016/0014-4827(87)90402-2
DO - 10.1016/0014-4827(87)90402-2
M3 - Article
C2 - 3653266
AN - SCOPUS:0023583151
SN - 0014-4827
VL - 172
SP - 439
EP - 449
JO - Experimental Cell Research
JF - Experimental Cell Research
IS - 2
ER -