Two-compartment model for rabbit skin organ culture |
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Authors: | A A J J L Rutten B G A G G Béquet-Passelecq H B W M Koëter |
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Institution: | (1) Department of Biological Toxicology, TNO-CIVO Toxicology and Nutrition Institute, P.O. Box 360, 3700 AJ Zeist, Netherlands |
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Abstract: | Summary A new method was developed for rabbit skin organ culture. In a two-compartment model, skin discs were cultured on a Millicell-HA
insert unit with a microporous membrane which allows transport of culture medium via the dermis into the epidermis, whereas
the epidermal side remains free of direct contact with culture medium. In this relatively simple two-compartment organ culture
model, rabbit skin could be cultured for 7 d in RPMI 1640 medium supplemented with fetal bovine serum, or for 2 d in RPMI
1640 medium supplemented with cofactors. The histomorphology and ultrastructure of 7-d cultured rabbit skin discs was essentially
similar to that of freshly isolated rabbit skin. Keratinocytes in the stratum basale continued to divide during organ culture.
The terminal differentiation of the epidermis continued in vitro as was found by the presence of keratohyalin granules, the
intact stratum corneum, and keratin expression. Furthermore, glucose consumption continued until culture Day 7, but thereafter
it declined rapidly. Concomitantly, degenerative changes were found. At the end of the 7-d culture period the distance between
single dermal collagen fibrils had increased as compared to noncultured skin. This model of skin organ cultures can be used
to study biological processes, dermal toxicity, and penetration and metabolism of xenobiotics in intact skin. Furthermore,
within certain limits, processes responsible for repair and regeneration of damaged skin can also be studied in this model
because the rabbit skin can be cultured for 7 d.
The present study was financially supported by grants of Duphar B. V. (Weesp, Netherlands), the European Community, and the
Dutch animal welfare organizations Samenwerkingsverband van de Nederlandse Vereniging tot Bescherming van Dieren en de Nederlandse
Bond tot Bestrijding van de Vivisectie, Anti-Vivisectie Stichting en Stichting Schoonheid Zonder Wreedheid. |
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Keywords: | skin culture organ culture dermal toxicity rabbit in vitro |
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