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. 1980 Jan;77(1):472–476. doi: 10.1073/pnas.77.1.472

Suspension culture of separated follicles consisting of differentiated thyroid epithelial cells

Lucio Nitsch 1,*, Seymour H Wollman 1,†
PMCID: PMC348293  PMID: 6244562

Abstract

We have prepared thyroid follicles in suspension culture to use as a model system in vitro for investigation of some properties of the thyroid gland. The follicles were free of endothelial cells, fibroblasts, and other nonepithelial cells. They were prepared by collagenase treatment of minced rat thyroid glands followed by differential filtration of the suspension through nylon meshes. Small clusters of principal thyroid epithelial cells were separated from large fragments and single cells. They were cultured in dilute suspension in Coon's modified F-12 medium in dishes coated with agarose to avoid having the cells attach to the dishes. By culture day 3, most of the clusters formed closed follicles containing a periodic acid-Schiff-positive colloid but without a basal lamina. Follicle walls contained an occasional C cell. The epithelium resembled that in the thyroid of a recently hypophysectomized rat, with normal polarity and organelle complement normal with respect to position and abundance, with basally located lysosomes, no pseudopods, and no colloid droplets. The cells were responsive to thyroid-stimulating hormone (thyrotropin) and to dibutyryl cyclic AMP. Thyroid-stimulating hormone at 10 munits/ml resulted in apical migration of lysosomes and formation of pseudopods and colloid droplets within 30 min; longer exposure resulted in depletion of luminal colloid. The results indicate that the suspended follicles resemble follicles in vivo with respect to morphology and responsiveness to thyroid-stimulating hormone in the absence of other cell types.

Keywords: ultrastructure, pseudopods, endocytosis, lysosomes

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Fayet G., Michel-Béchet M., Lissitzky S. Thyrotrophin-induced aggregation and reorganization into follicles of isolated porcine-thyroid cells in culture. 2. Ultrastructural studies. Eur J Biochem. 1971 Dec 22;24(1):100–111. doi: 10.1111/j.1432-1033.1971.tb19659.x. [DOI] [PubMed] [Google Scholar]
  2. Fayet G., Pacheco H., Tixier R. Sur la reassociation in vitro des cellules isolees de thyrïde de porc et la biosynthése de la thyroglobuline. I. Conditions pour l'induction des reassociations cellulaires par la thyreostimuline. Bull Soc Chim Biol (Paris) 1970 Apr 17;52(3):299–306. [PubMed] [Google Scholar]
  3. Hutchings S. E., Sato G. H. Growth and maintenance of HeLa cells in serum-free medium supplemented with hormones. Proc Natl Acad Sci U S A. 1978 Feb;75(2):901–904. doi: 10.1073/pnas.75.2.901. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Isler H., Sarkar K., Thompson B., Tonkin R. The architecture of the thyroid gland: a 3-dimensional investigation. Anat Rec. 1968 Jul;161(3):325–335. doi: 10.1002/ar.1091610305. [DOI] [PubMed] [Google Scholar]
  5. KERKOF P. R., LONG P. J., CHAIKOFF I. L. IN VITRO EFFECTS OF THYROTROPIC HORMONE. I. ON THE PATTERN OF ORGANIZATION OF MONOLAYER CULTURES OF ISOLATED SHEEP THYROID GLAND CELLS. Endocrinology. 1964 Feb;74:170–179. doi: 10.1210/endo-74-2-170. [DOI] [PubMed] [Google Scholar]
  6. Lissitzky S., Fayet G., Giraud A., Verrier B., Torresani J. Thyrotrophin-induced aggregation and reorganization into follicles of isolated porcine-thyroid cells. 1. Mechanism of action of thyrotrophin and metabolic properties. Eur J Biochem. 1971 Dec 22;24(1):88–99. doi: 10.1111/j.1432-1033.1971.tb19658.x. [DOI] [PubMed] [Google Scholar]
  7. Pastan I., Wollman S. H. Colloid droplet formation in dog thyroid in vitro. Induction by dibutyrl cyclic-AMP. J Cell Biol. 1967 Oct;35(1):262–266. doi: 10.1083/jcb.35.1.262. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Rapoport B. Dog thyroid cells in monolayer tissue culture: adenosine 3', 5'-cyclic monophosphate response to thyrotropic hormone. Endocrinology. 1976 May;98(5):1189–1197. doi: 10.1210/endo-98-5-1189. [DOI] [PubMed] [Google Scholar]
  9. Studer H., Forster R., Conti A., Kohler H., Haeberli A., Engler H. Transformation of normal follicles into thyrotropin-refractory "cold" follicles in the aging mouse thyroid gland. Endocrinology. 1978 May;102(5):1576–1586. doi: 10.1210/endo-102-5-1576. [DOI] [PubMed] [Google Scholar]
  10. Tice L. W. Effects of hypophysectomy and TSH replacement on the ultrastructural localization of thyroperoxidase. Endocrinology. 1974 Aug;95(2):421–433. doi: 10.1210/endo-95-2-421. [DOI] [PubMed] [Google Scholar]
  11. WOLLMAN S. H., SPICER S. S., BURSTONE M. S. LOCALIZATION OF ESTERASE AND ACID PHOSPHATASE IN GRANULES AND COLLOID DROPLETS IN RAT THYROID EPITHELIUM. J Cell Biol. 1964 May;21:191–201. doi: 10.1083/jcb.21.2.191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Wetzel B. K., Spicer S. S., Wollman S. H. Changes in fine structure and acid phosphatase localization in rat thyroid cells following thyrotropin administration. J Cell Biol. 1965 Jun;25(3):593–618. doi: 10.1083/jcb.25.3.593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Winand R. J., Kohn L. D. Thyrotropin effects on thyroid cells in culture. Effects of trypsin on the thyrotropin receptor and on thyrotropin-mediated cyclic 3':5'-AMP changes. J Biol Chem. 1975 Aug 25;250(16):6534–6540. [PubMed] [Google Scholar]
  14. Zeligs J. D., Wollman S. H. Pseudopod behavior in hyperplastic thyroid follicles in vivo. J Ultrastruct Res. 1977 Jul;60(1):99–105. doi: 10.1016/s0022-5320(77)80046-4. [DOI] [PubMed] [Google Scholar]

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