Reconstitution of human epidermis in vitro is accompanied by transient activation of basal keratinocyte spreading

Frederick Grinnell, Ken Ichi Toda, Cheryl Lamke-Seymour

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Abstract

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.

Original languageEnglish (US)
Pages (from-to)439-449
Number of pages11
JournalExperimental Cell Research
Volume172
Issue number2
DOIs
StatePublished - 1987

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Keratinocytes
Epidermis
Skin
Basement Membrane
Fibronectins
Hemidesmosomes
Electrons
Staining and Labeling
Desmosomes
Collagen Type IV
Antibodies
Serum-Free Culture Media
Laminin
Indirect Fluorescent Antibody Technique
Cadaver
Collagen
In Vitro Techniques

ASJC Scopus subject areas

  • Cell Biology

Cite this

Reconstitution of human epidermis in vitro is accompanied by transient activation of basal keratinocyte spreading. / Grinnell, Frederick; Toda, Ken Ichi; Lamke-Seymour, Cheryl.

In: Experimental Cell Research, Vol. 172, No. 2, 1987, p. 439-449.

Research output: Contribution to journalArticle

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