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Title: Electrophysiological studies on primary cultures of skin cells
Author: Dove, N. S.
Awarding Body: University of Cambridge
Current Institution: University of Cambridge
Date of Award: 1997
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To model sweat gland fluid secretion in vitro, human apocrine glands were isolated from axillary skin and their secretory coil cells were cultured and mounted in an Ussing chamber. Cultured apocrine coil cells exhibited an amiloride-sensitive inward resting short circuit current, suggesting an absorptive phenotype. However, evidence of a chloride-secretory phenotype was also observed: transient inward currents were recorded in response to autonomic secretagogues and inflammatory mediators, and these were not abolished by amiloride, but they were reduced by half in chloride-free buffer. This may indicate a role for chloride secretion in mediating the stimulated currents. Further, sustained inward currents were recorded in 58% of tissues which were attenuated by frusemide, implying the involvement of the Na+/K+/Cl- transporter. Assuming that secondary active chloride transport is responsible for aqueous fluid secretion in the apocrine sweat gland, these data are compatible with some maintenance of an in vivo phenotype in culture. To study the cellular determinants of eccrine sweat gland differentiation, eccrine sweat gland-associated fibroblasts were cultured and compared with derminal fibroblasts from the same subject, demonstrated a different pattern of outgrowth and proliferation. Therefore these cells may represent a novel fibroblast subtype. To improve the differentiation of eccrine sweat gland coil cells in vitro, they were co-cultured with eccrine fibroblasts. Eccrine sweat gland cells grown on a Transwell as a control demonstrated apparent dedifferentiation to a reabsorptive phenotype, indicated by the amiloride-sensitivity of their resting and agonist-stimulated inward short circuit currents. Eccrine cells co-cultured with eccrine fibroblasts, however, demonstrated agonist-stimulated outward currents in the presence of amiloride.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available