Use of a Serum-Free Epidermal Culture Model to Show Deleterious Effects of Epidermal Growth Factor on Morphogenesis and Differentiation

The presence of serum has limited the utility of many culture models for the study of cytokine effects because Its complexity and variability can confound the interpretation of data. In the present study, a serum-free skin co-culture model was used to investigate the effect of exogenous epidermal gr...

Full description

Saved in:
Bibliographic Details
Published in:Journal of investigative dermatology 1995-01, Vol.104 (1), p.107-112
Main Authors: Chen, Chih-Shan J., Lavker, Robert M., Rodeck, Ulrich, Risse, Barbara, Jensen, Pamela J.
Format: Article
Language:English
Subjects:
Air
Antigens, Differentiation - analysis
Cell Differentiation - drug effects
Cell Division - drug effects
Cells, Cultured
Culture Media, Serum-Free - pharmacology
Epithelium - ultrastructure
ErbB Receptors - analysis
Fibroblasts - cytology
Frozen Sections
Humans
Immunohistochemistry
keratinocyte
Keratinocytes - cytology
proliferation
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:The presence of serum has limited the utility of many culture models for the study of cytokine effects because Its complexity and variability can confound the interpretation of data. In the present study, a serum-free skin co-culture model was used to investigate the effect of exogenous epidermal growth factor (EGF) on epidermal proliferation and differentiation, Human keratinocytes cultured on collagen rafts at the air-liquid interface produced a well-differentiated epithelium that resembled normal epidermis. Keratin filaments, membrane-coating granules, and keratohyalin granules were all observed. Epidermal differentiation markers keratin K1/K10, involucrin, and transglutaminase were localized in most of the supra-basal layers, whereas profilaggrin/filaggrin was confined to the granular layers and stratum corneum, In the continual presence of 10–20 ng/mL EGF, the epidermis was less organized, thinner, and less proliferative. EGF also depressed several indicators of differentiation: The number of keratohyalin granules and membrane-coating granules was greatly decreased; antigen expression of profilaggrin/filaggrin appeared diminished by immunocytochemical staining; frequent nuclear retention was noted in the relatively thickened stratum corneum-like layers. As detected by immunohistochemical staining, the expression of EGF receptor in the epidermis was reduced by exogenous EGF. These data illustrate that EGF cannot be considered a simple mitogen. Our findings also underscore tile importance of using sophisticated culture models to assess complex cytokine effects that may be dependent on the architecture of a differentiating epidermis.
ISSN:0022-202X
1523-1747
DOI:10.1111/1523-1747.ep12613595