Iodine metabolism of dispersed thyroid cells obtained by trypsinization of sheep thyroid glands

Tong, Winton; Kerkof, Paul R.; Chaikoff, I.L.

doi:10.1016/0006-3002(62)90365-7

Cited by 82 publications

(25 citation statements)

References 18 publications

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“…Trypsin or collagenase have usually been used for the isolation of thyroid cells (Tong et al, 1962 ;Nitsch and Wollman, 1980). We showed that dispase was also an excellent tool for isolation of thyroid cells.…”

Section: Discussionmentioning

confidence: 99%

An ecto-ATPase of thyroidal cell membrane.

1983

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The isolated cells were obtained from hog thyroid glands treated with dispase. More than 95 % of the cells obtained were intact and viable immediately after preparation, and the cell viability did not change during incubation in the experimental conditions. ATP added to the external medium of whole cell suspensions was hydrolyzed in the presence of various divalent cations, especially Mg, and the rate of hydrolysis of ATP was not significantly different between the Mg-ion system and the completed ion system (Mg+ Na +K). When whole cell suspensions were disrupted with homogenizer, the hydrolysis of ATP was markedley increased by adding Na plus K. But there was no difference in the Mg-ion system between cell homogenates and whole cell suspensions.ADP, AMP and adenosine as reaction products were found in the reaction mixture which resulted from the hydrolysis of ATP by whole cell suspensions.Our data suggest that Mg-ATPase in the thyroidal isolated cells is an ectoenzyme whose active site(s) are exposed to the external surface of plasma membrane, and that ATP is finally hydrolyzed to adenosine via ADP and AMP by the enzyme(s).

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Section: Discussionmentioning

confidence: 99%

An ecto-ATPase of thyroidal cell membrane.

1983

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“…Isolated thyroid cells from various species, including human beings, can be maintained in culture for a limited period of time (1)(2)(3)(4)(5)(6)(7)(8)(9)(10). The addition of thyrotropin (TSH) to the cultures at the time of explantation makes the cells organize into follicular structures (3)(4)(5)(6)(7) that maintain the capacity to concentrate iodide and synthesize iodinated thyroglobulin (7,11).…”

mentioning

confidence: 99%

Thyrotropin is not a growth factor for human thyroid cells in culture

Westermark

Karlsson

Wålinder

1979

Proc. Natl. Acad. Sci. U.S.A.

107

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Thyroid cells, obtained from both normal human tissue and benign nodular goiter, were cultured and maintained in vitro in 4-18 passages. Cultures with confluent cells accumulated cyclic AMP (10-150 times the basal amount) upon addition of bovine thyrotropin (100 milliunits/ml), indicating that the cells in culture maintained a thyrotropin-sensitive adenylate cyclase system. Addition of high doses of thyrotropin also induced a characteristic and reversible change in the morphology of the cells.The effect of thyrotropin on cell growth was studied in short- (HtTh 14). The tissue was fragmented with a scalpel, suspended in phosphate-buffered saline containing 1 mg of collagenase per ml, and incubated in a 37°C rocking waterbath for 60 min. Cells were further liberated from the fragments by pipetting. Large fragments were allowed to sediment and the supernatant was decanted and centrifuged (200 X g for 10 min). The pellets were suspended in Eagle's minimum essential medium (14) supplemented with 10% fetal calf serum, 100 units of penicillin, 1.25 ,gg of amphotericin B, and 50 ,ug of streptomycin per ml. The cells were incubated in 50-mm Nunc petri dishes (Nunc, Roskilde, Denmark). About 20 dishes were initiated from 1 g of wet weight tissue. The cultures were incubated at 37°C in a humidified atmosphere containing 5% CO2. Primary confluent monolayers were usually formed within 7-14 days. The cultures were then subcultivated at 1:2 split ratio. Cells were detached after incubation in phosphate-buffered saline with 0.02% EDTA for 5 min followed by a film of phosphate-buffered saline with 0.25% trypsin for about 10 min. Cultures of normal human skin fibroblasts and human glial cells were initiated and maintained as described (15,16 Abbreviation: TSH, thyrotropin.

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“…DISCUSSION Thyrocyte suspensions have been prepared previously by trypsinization of thyroid tissue (cf. Pulvertaft et al 1959;Tong et al 1962;Hung 8c Winskip 1964;Rodesch 8c Dumont 1967). When prepared from porcine thyroid, the cells did not respond to TSH except after cultivation for 12 h (Lissitzky et al 1973).…”

Section: Preparation Of Hormone-responsive Thyroid Cell Suspensionsmentioning

confidence: 97%

Isolation of TSH-Responsive Human Thyrocytes-Analysis of Various Thyroid Tumours

Breustedt

Nolde

Schumacher

et al. 1977

Acta Endocrinologica

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A method for the isolation of hormone-responsive thyrocyte suspensions from human tumours is described.Cells from normal and adenomatous thyroid tissue responded to thyroid stimulating hormone (TSH) by a concentration-dependent increase in cAMP. Bovine TSH usually produced half-maximal elevation of cAMP levels at concentrations between 2 and 4 mU/ml. Human TSH caused hal f\x=req-\ maximal stimulation at 3\p=n-\8 times lower concentrations indicating considerable species specificity of the human TSH receptor. Other hormones were without influence except prostaglandin E1 (PGE1) which evoked moderate elevations of the cAMP levels.Kinetic analyses of various thyroid tumours revealed marked differences in TSH induced increases in the intracellular cAMP levels with respect to the time course as well as to the extent of the maximal response. Theophylline, which by itself had a rather small effect on basal cAMP levels, acted synergistically with TSH in most cases, leading to values up to 20 times above the control values. The kinetics also indicated the existence in stimulated cells of two cAMP pools exhibiting different susceptibility to phosphodiesterase. Cells from adenomatous goitres with hyperplasia exhibited significantly lower basal cAMP levels than thyrocytes from the other goitres and from normal tissue. Other classes of thyroid tumours also showed characteristic differences with regard to basal cAMP and degree of stimulation by TSH and theophylline.

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Iodine metabolism of dispersed thyroid cells obtained by trypsinization of sheep thyroid glands

Cited by 82 publications

References 18 publications

An ecto-ATPase of thyroidal cell membrane.

An ecto-ATPase of thyroidal cell membrane.

Thyrotropin is not a growth factor for human thyroid cells in culture

Isolation of TSH-Responsive Human Thyrocytes-Analysis of Various Thyroid Tumours

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