Culture of sweat gland epithelial cells from normal individuals and patients with cystic fibrosis

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Abstract
Recent electrophysiological and pharmacological studies have confirmed previous clinical evidence that the gene defect in cystic fibrosis is strongly expressed in the sweat gland. This has provided a major impetus to efforts to culture the cells of this tissue in order to provide a source of experimental material for molecular studies. Toward this end, eccrine sweat glands were isolated from collagenase treated skin specimens and the secretory coil and the reabsorptive duct separated. Segments of each portion of the gland were transferred to a plastic or collagen substrate and covered with serum-containing or serum-free defined growth media. Epithelial cell outgrowth took place in both media but fibroblast overgrowth occurred in the presence of serum at concentrations as low as 1%. In serum-free medium both secretory and reabsorptive cells formed tightly joined epithelial sheets, first as monolayers and later as multilayers consisting of at least six cell layers. Growth continued for approximately fifteen generations each of about two and a half days. Remarkably large domes or hemicysts with diameters as great as two cm were formed, apparently attesting to the retention of the capacity of the cells to actively transport ions and water. Ultrastructurally, cells which grew out from the secretory coil ressembled the fluid secreting clear cells; neither dark cells nor myoepithelial cells were propagated.