Abstract

Human skin organ cultures were established from 2-mm punch biopsies and incubated under serum-free conditions in basal medium containing either 0.15 or 1.4 mM extracellular Ca2+. Organ cultures were treated with concentrations of sodium lauryl sulfate (SLS) that had previously been shown to support growth of human epidermal keratinocytes and human dermal fibroblasts in monolayer culture. Epidermal growth factor (EGF), alone and in combination with insulin and bovine pituitary extract, fetal bovine serum and all-trans retinoic acid (RA) were also examined for comparative purposes. The addition of SLS to culture medium containing low extracellular Ca2+ had no effect on the viability or histological appearance of the organ-cultured skin. Complete degeneration of the tissue occurred in the presence of SLS just as it did under control conditions. When SLS was added to culture medium containing high extracellular Ca2+, the basal layer of keratinocytes was much thinner than under control conditions. When EGF or EGF in combination with insulin and pituitary extract were utilized in place of SLS, identical results were obtained. That is, there was no preservation of the basal epithelial layer in the presence of low-Ca2+ culture medium and in the presence of high-Ca2+ culture medium, the basal layer was thinner than in control tissue. Virtually identical results were also obtained in medium containing 10% fetal bovine serum. In contrast, when RA was included in low-Ca2+ culture medium, the basal epithelium was maintained in a viable, histologically healthy condition. However, normal epithelial differentiation did not occur and the upper layers of the epidermis separated from the basal cells.(ABSTRACT TRUNCATED AT 250 WORDS)

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