The primary culture of epithelia from the secretory coil and collecting duct of normal human and cystic fibrotic eccrine sweat glands

1986 ◽  
Vol 83 (1) ◽  
pp. 103-118
Author(s):  
C.M. Lee ◽  
F. Carpenter ◽  
T. Coaker ◽  
T. Kealey
Keyword(s):  
Ion Transport ◽  
Collecting Duct ◽  
Serum Insulin ◽  
Primary Cultures ◽  
Calf Serum ◽  
Sweat Glands ◽  
Foetal Calf Serum ◽  
Secretory Coil ◽  
Eccrine Sweat Glands ◽  
Eccrine Sweat

Isolated human eccrine sweat glands have been microdissected into their secretory and reabsorptive components. Complete separation of these epithelia was confirmed by differential uptake of Neutral Red stain by an intermediate section of gland containing the junction between the secretory coil and the collecting duct. Primary cultures were obtained from explants of both tissues in medium RPMI-1640 or Williams E supplemented with foetal calf serum, insulin, transferrin, epidermal growth factor and hydrocortisone. The cells in the initial coil cultures had an elongated morphology while those of ductal origin were polyhedral. After 10 days both cultures were composed of polyhedral cells of varying diameter. All these morphological types were of epithelial lineage, as demonstrated by the binding of a monoclonal antibody to cytokeratin, the intermediate filament specific for epithelial cells. Outgrowth from both secretory and reabsorptive epithelia were multilayered, with plentiful desmosomal connections and an underlying basal lamina. Ultrastructural features typical of the epithelial cell types present in intact eccrine sweat glands were absent in a high proportion of the proliferating cells but domes, indicative of transepithelial active ion transport, were present in dense cultures from the reabsorptive duct. Outgrowth was also obtained from the secretory and reabsorptive epithelia of sweat glands from two cystic fibrotic patients. Since the most characteristic malfunction of cystic fibrosis is the impaired ion transport in the eccrine sweat gland, the availability of cultured epithelia should provide a useful model for study of the disease.

Freeze-Fracture Examination of Tight Junctions of Human Eccrine Sweat Glands

1980 ◽  
Vol 38 ◽  
pp. 634-635
Author(s):  
J. V. Briggman ◽  
J. Bigelow ◽  
H. Bank ◽  
S. S. Spicer
Keyword(s):  
Cystic Fibrosis ◽  
Freeze Fracture ◽  
Sweat Glands ◽  
Hypertonic Solution ◽  
Dark Cells ◽  
Clear Cells ◽  
Junctional Complexes ◽  
Secretory Coil ◽  
Eccrine Sweat Glands ◽  
Eccrine Sweat

The prevalence of strands shown by freeze-fracture in the zonula occludens of junctional complexes is thought to correspond closely with the transepi-thelial electrical resistance and with the tightness of the junction and its obstruction to paracellular flow.1 The complexity of the network of junc¬tional complex strands does not appear invariably related to the degree of tightness of the junction, however, as rabbit ileal junctions have a complex network of strands and are permeable to lanthanum. In human eccrine sweat glands the extent of paracellular relative to transcellular flow remains unknown, both for secretion of the isotonic precursor fluid by the coil and for resorption of a hypertonic solution by the duct. The studies reported here undertook, therefore, to determine with the freeze-fracture technique the complexity of the network of ridges in the junctional complexes between cells in the secretory coil and the sweat ducts. Glands from a patient with cystic fibrosis were also examined because an alteration in junctional strands could underlie the decreased Na+ resorption by sweat ducts in this disease. Freeze-fracture replicas were prepared by standard procedures on isolated coil and duct segments of human sweat glands. Junctional complexes between clear cells, between dark cells and between clear and dark cells on the main lumen, and between clear cells on intercellular canaliculi of the coil con¬tained abundant anastomosing closely spaced strands averaging 6.4 + 0.7 (mean + SE) and 9.0 +0.5 (Fig. 1) per complex, respectively. Thus, the junctions in the intercellular canaliculi of the coil appeared comparable in complexity to those of tight epithlia. Occasional junctions exhibited, in addition, 2 to 5 widely spaced anastomosing strands in a very close network basal to the compact network. The fewer junctional complexes observed thus far between the superficial duct cells consisted on the average of 6 strands arranged in a close network and 1 to 4 underlying strands that lay widely separated from one another (Fig. 2). The duct epitelium would, thus, be judged slightly more "leaky" than the coil. Infrequent junctional complexes observed to date in the secretory coil segment of a cystic fibrosis specimen disclosed rela¬tively few closely crowded strands.

NCL-SG3: a human eccrine sweat gland cell line that retains the capacity for transepithelial ion transport

1989 ◽  
Vol 92 (2) ◽  
pp. 241-249
Author(s):  
C.M. Lee ◽  
J. Dessi
Keyword(s):  
Epithelial Cell ◽  
Ion Transport ◽  
Cell Line ◽  
Intermediate Filament ◽  
Primary Cultures ◽  
Simian Virus 40 ◽  
T Antigen ◽  
Sweat Glands ◽  
Epithelial Cell Line ◽  
Eccrine Sweat

An ion-transporting human epithelial cell line, NCL-SG3, has been established by simian virus 40 (SV40) infection of primary cultures from eccrine sweat glands. The line has been passaged 38 times (over 100 population doublings), has an aneuploid karyotype but has not undergone any ‘crisis’. The cells have retained epithelial morphology and expression of cytokeratin, the intermediate filament characteristic of epithelial cells. Approximately 85% of the population shows at least weak co-expression of vimentin, an intermediate filament associated with mesenchymal and some other non-epithelial cell types in vivo. In addition, SV40 large T-antigen is present, in a predominantly nuclear localization. Electrically resistant cell sheets are formed on dialysis tubing and cellulose-ester permeable supports. Electrogenic ion transport can be stimulated by the beta-adrenergic agonist isoproterenol (10(−6) M) and by lysylbradykinin (10(−7) M) but not by the cholinergic agonist carbachol at 10(−6) M).

Biochemical and ultrastructural studies of human eccrine sweat glands isolated by shearing and maintained for seven days

1984 ◽  
Vol 72 (1) ◽  
pp. 259-274
Author(s):  
C.M. Lee ◽  
C.J. Jones ◽  
T. Kealey
Keyword(s):  
Cyclic Amp ◽  
Cyclic Gmp ◽  
Collecting Duct ◽  
Energy Charge ◽  
Light And Electron Microscopy ◽  
Sweat Glands ◽  
Ultrastructural Studies ◽  
Leucine Uptake ◽  
Eccrine Sweat Glands ◽  
Eccrine Sweat

A new method of isolating human eccrine sweat glands by the repeated dissection of skin biopsies with scissors is described. The success of the technique is attributed to a potential line of weakness between the investing capsule and the surrounding connective tissue, which parts under shear forces. The yield is 20–50 glands per biopsy (5 cm X 0.5 cm). The glands are judged to be viable by: (i) light and electron microscopy; (ii) ATP, ADP and AMP contents of 81.0 +/− 12.7, 13.8 +/− 3.3 and 3.8 +/− 1.0 pmol/gland, respectively (mean +/− S.E.M.), which gave an energy charge of 0.90; (iii) the 28-fold rise in cyclic GMP content and the sevenfold rise in cyclic AMP content effected by treatment for 2 min with 10(−5) M-acetylcholine and for 10 min with 10(−5) M-isoprenaline, respectively; (iv) the rate of [3H]leucine uptake into protein; and (v) the concentration of Neutral Red by the collecting duct. Glands were maintained for 7 days on polycarbonate filters floating on RPMI 1640 tissue-culture medium. After this time the ATP, ADP and AMP contents were 63.2 +/− 7.3, 8.5 +/− 2.2 and 3.5 +/− 0.8 pmol/gland, respectively (mean +/− S.E.M.), which gave an energy charge of 0.90. During maintenance a dilatation of the intercellular spaces developed in both secretory coil and collecting duct. Following maintenance there was a significant rise in the rate of [3H]leucine uptake into protein. Maintained glands demonstrated a fivefold greater accumulation of cyclic AMP in response to isoprenaline than did freshly isolated glands, but there was no comparable maintenance hypersensitivity of cyclic GMP to acetylcholine. This pattern of adrenergic, but not cholinergic, maintenance hypersensitivity matches the known lack of denervation hypersensitivity of human eccrine sweat glands to acetylcholine in vivo.

scholarly journals Training of Eccrine Sweat Glands in Paraplegics

1970 ◽  
Vol 54 (3) ◽  
pp. 229-232 ◽  
Author(s):  
Becky B. Johnson ◽  
Robert E. Johnson
Keyword(s):  
Sweat Glands ◽  
Eccrine Sweat Glands ◽  
Eccrine Sweat

Tufted angioma associated with hyperplasia of eccrine sweat glands

2017 ◽  
Vol 42 (5) ◽  
pp. 548-550
Author(s):  
Y. Saito ◽  
Y. Shimomura ◽  
R. Abe
Keyword(s):  
Sweat Glands ◽  
Tufted Angioma ◽  
Eccrine Sweat Glands ◽  
Eccrine Sweat

Pharmacologic responsiveness of isolated single eccrine sweat glands

1981 ◽  
Vol 240 (1) ◽  
pp. R44-R51 ◽  
Author(s):  
K. Sato ◽  
F. Sato
Keyword(s):  
Dose Response ◽  
Dissociation Constants ◽  
Sweat Rate ◽  
Sweat Glands ◽  
Adrenergic Agonist ◽  
Beta Adrenergic ◽  
Adrenergic Agents ◽  
Eccrine Sweat Glands ◽  
Eccrine Sweat

Pharmacologic responsiveness of the eccrine sweat gland has never been studied under well-defined in vitro experimental conditions. Using isolated cannulated single monkey palm eccrine sweat glands, the dose response to both cholinergic and alpha- and beta-adrenergic agents and the effects of various antagonists on agonists were studied. The maximal sweat rate was highest after stimulation with cholinergic agonists, was lower with the beta-adrenergic agonist, and was least with the alpha-adrenergic agonist. Each secretory response was inhibited by its specific antagonist. Attempts to demonstrate the spare receptor, if any, by means of preincubation of the glands with N-(2-chlorethyl)dibenzylamine (Dibenamine) were unsuccessful. From the hyperbolic dose-response curves the values for KA and KB, dissociation constants for agonists and antagonists, respectively, were thus tentatively estimated according to Clark's classical receptor theory. Schild plots for each agonist-antagonist interaction produced straight lines with slopes of near unity, indicating the adequacy of the methodology. It was concluded that the isolated eccrine sweat glands retain their pharmacologic viability in vitro and show responsiveness to cholinergic as well as both alpha- and beta-adrenergic stimulations.

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