Alginate Gel Culture Allows the Retention of Extracellular Matrix and Follicular Structure of Rat Thyroid Tissue But Does Not Lead to the Formation of Follicles by FRTL-5 Cells

Burgi-Saville, M. E.; Reut, Barbara; Gerber, H; Peter, H J; Paulsson, Mats; Kaempf, Jacqueline; Simon, Franziska; Marti, Ulrich; Gerber, H.; Bürgi, U

doi:10.1089/thy.1998.8.1147

Cited by 14 publications

(15 citation statements)

References 16 publications

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“…In contrast, FRTL-5 cells treated under the same conditions were not observed to make follicles. This observation is consistent with those of a recent study attempting to induce follicles in FRTL-5 cells using a different approach (Burgi-Saville et al 1998).…”

Section: Follicle Reconstitution By Long-term Cultured Normal Human Tsupporting

confidence: 93%

Induction of follicle formation in long-term cultured normal human thyroid cells treated with thyrotropin stimulates iodide uptake but not sodium/iodide symporter messenger RNA and protein expression

Kogai¹,

Curcio²,

Hyman³

et al. 2000

Journal of Endocrinology

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Iodide uptake by the sodium/iodide symporter (NIS) in thyrocytes is essential for thyroid hormone production. Reduced NIS activity has been reported in thyroid diseases, including thyroid cancer and congenital hypothyroidism. The study of iodide uptake in thyrocytes has been limited by the availability of appropriate in vitro models. A new culture technique was recently developed that allows normal human thyroid primary culture cells to grow as monolayer cells and express differentiated functions for more than 3 months. We used this technique to study the effect of follicle formation and TSH on iodide uptake in these cells. Iodide uptake by the cells grown in monolayer was very low. Follicle formation was induced from monolayer cells, and electron micrographs demonstrated cell polarity in the follicles. No significant increase in iodide uptake was observed after TSH treatment of cells in monolayer or when follicle formation was induced without TSH. TSH stimulation of follicles, however, significantly increased iodide uptake (4·4-fold; P<0·001). Compared with iodide uptake in monolayers, the combination of follicle formation and TSH treatment stimulated iodide uptake synergistically to 12·0-fold (P<0·001). NIS messenger RNA (mRNA) and protein levels were almost the same in both monolayer cells and follicles. TSH treatment of monolayers and follicles produced significant (P<0·05) stimulation of mRNA (4·8-and 4·3-fold respectively) and protein (6·8-and 4·9-fold respectively). TSH stimulated NIS protein levels in both monolayer and follicles, however, stimulation of functional iodide uptake was only seen with TSH stimulation of follicles. The function of NIS may involve post-transcriptional events, such as intracellular sorting, membrane localization of NIS or another NIS regulatory factor. Polarized functions, such as iodide efflux into follicular lumina, may also contribute to the increased iodide concentration after follicle formation.

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Section: Follicle Reconstitution By Long-term Cultured Normal Human Tsupporting

confidence: 93%

Induction of follicle formation in long-term cultured normal human thyroid cells treated with thyrotropin stimulates iodide uptake but not sodium/iodide symporter messenger RNA and protein expression

Kogai¹,

Curcio²,

Hyman³

et al. 2000

Journal of Endocrinology

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“…Synthesis of laminin, CoIV and fibronectin has been demonstrated in the thyroid epithelial cell line FRTL5 (Ambesi-Impiombato et al 1980, Ambesi-Impiombato & Perrild 1989 and also the deposition of the extracellular material at the basolateral surface of the cells and its organization into a BM (Garbi et al 1988). We have recently demonstrated the maintenance of an ECM containing fibronectin, CoIV, laminin and perlecan both in FRTL5 cells and in primary cultures of rat thyroid cells (Bürgi-Saville et al 1998) and in primary cultures of normal human thyroid tissue, also in alginate beads (Bürgi-Saville et al 1997).…”

Section: Introductionmentioning

confidence: 74%

“…In order to study in vitro thyroid differentiation, function and proliferation, various cell culture systems have been tested: monolayers (Ambesi-Impiombato et al 1980, Ambesi-Impiombato & Perrild 1989, Derwahl et al 1990, Gerber et al 1991a,b, Peter et al 1991b, Dumont et al 1992, three-dimensional systems using collagen gels (Yang et al 1979, Garbi & Wollman 1982, Garbi et al 1984, Peter et al 1987, 1991a-c, Toda & Sugihara 1990, Toda et al 1993, agarose gels (Kimura et al 1980, Benya & Shaffer 1982, Aydelotte et al 1986a,b, Bruckner et al 1989) and alginate beads (Kupchick et al 1983, Guo et al 1989, Häuselmann et al 1994, Tognella et al 1995, Glaser et al 1996, Bürgi-Saville et al 1997, 1998. Alginate has several advantages in comparison with the other systems.…”

Section: Introductionmentioning

confidence: 99%

“…So far, much less but nevertheless fascinating research has been done on the structure and metabolism of ECM of thyroid tissue (Garbi & Wollman 1982, Alquier et al 1983, Garbi et al 1984, Wadeleux et al 1985, Demeure et al 1992, Prabakaran et al 1993, Vitale et al 1993, 1994, Andre et al 1994, Lemansky et al 1994, Tognella et al 1995, Arvan et al 1997, Bürgi-Saville et al 1997, 1998. Ultrastructural studies indicate that BM formation or maintenance can be achieved in cultures of thyroid cells (Alquier et al 1983, Garbi & Wollman 1982, Garbi et al 1988.…”

Section: Introductionmentioning

confidence: 99%

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Follicle-forming cat thyroid cell lines synthesizing extracellular matrix and basal membrane components: a new tool for the study of thyroidal morphogenesis

Tognella

Marti²,

Peter³

et al. 1999

Journal of Endocrinology

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Interactions between follicular epithelial cells and extracellular matrix (ECM) are supposed to play an important role in the development and maintenance of thyroid tissue architecture. In the present study we have therefore investigated the synthesis of ECM components by a feline thyroid cell line which is able to form follicle-like structures in vitro, and also in v-ras-transfected and controltransfected sublines. Transfections were performed by lipofection with pZSR (viral Harvey ras gene; neo) and pSV2-neo (control, neo only) plasmids. We have adapted a semisolid culture system composed exclusively of polymerized alginate and therefore devoid of ECM components. Feline cells embedded in alginate gels as single cells and cultured for up to 90 days formed cell clusters within 10 days. Follicle-like structures were formed in the original cell lines and also in the v-ras-and controltransfected cells. Differences in proliferation rates were observed, the v-ras-transfected cells growing up to two to three times faster than the non-transfected cells. Immunostaining was done using rabbit first antibodies directed against mouse collagen IV, human fibronectin, laminin (tumor Engelbreth-Holm-Swarm laminin), perlecan and other ECM components. For comparison, immunostaining was also performed on cryosections of nodular goiters of six hyperthyroid cats. The cell lines and their transfected clones stained strongly positive for collagen IV and fibronectin, and positively but less strongly for laminin and perlecan. The cat goiter tissue stained positively for collagen IV, laminin, perlecan, and fibronectin, and positive staining for S-laminin (containing the 2-chain) was seen in blood vessel walls in this tissue.In conclusion, cat cell lines grow three-dimensionally in alginate beads over several weeks, they form follicle-like structures and express the same ECM components as the native cat goiter tissue. Transfection with v-ras does increase proliferation rate, but does not fundamentally alter formation of follicle-like structures and ECM expression. Alginate gel culture is a promising new tool for the study of follicular morphogenesis, polarity, the expression pattern of ECM components and of the interaction between thyrocytes and ECM. It avoids interference caused by gels composed of ECM components.

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“…In a further report, we were able to show the ability of the alginate gel culture system to retain ECM and follicular structure of rat thyroid tissue. In addition, iodine organification was maintained in the cultured organoids (14).…”

Section: Introductionmentioning

confidence: 99%

Inhibition of Iodine Organification and Regulation of Follicular Size in Rat Thyroid Tissue In Vitro

Glaser

Marti

Burgi-Saville

et al. 1999

ENDO

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The factors mediating the accumulation of thyroglobulin are of great importance to the understanding of the pathogenesis of human and experimentally induced colloid goiters. To elucidate further the underlying cellular mechanism, thyroid fragments from newborn rats were incorporated into semisolid alginate beads and were cultured as three-dimensional organoids for up to 21 d. In five parallel cultures, the medium contained either no supplements (group A), Nal (group B), thyroid-stimulating hormone (TSH) (group C), Nal plus TSH in the same concentrations as B and C (group D), or Nal and TSH (as in group D) plus methimazole (MMI, group E). The thyroid organoids maintained morphological integrity, functional activity, and ability to proliferate in vitro. Addition of iodine to the cultures significantly increased mean (+/-SEM) follicular diameters from 19.5 +/- 0.7 microm in controls to 33.9 +/- 2.2 microm (p < 0.0001) when NaI was added alone (group B), and 30.4 +/- 1.7 microm (p < 0.0001) when combined with TSH (group D). The effect of NaI on follicular size was abolished by MMI (group E, follicular diameter 23.5 +/- 1.3 microm). The results presented support the recent finding, using a rat colloid goiter model, that not only TSH but also iodine organification or its inhibition are important factors in modulating follicular morphology.

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Alginate Gel Culture Allows the Retention of Extracellular Matrix and Follicular Structure of Rat Thyroid Tissue But Does Not Lead to the Formation of Follicles by FRTL-5 Cells

Cited by 14 publications

References 16 publications

Induction of follicle formation in long-term cultured normal human thyroid cells treated with thyrotropin stimulates iodide uptake but not sodium/iodide symporter messenger RNA and protein expression

Induction of follicle formation in long-term cultured normal human thyroid cells treated with thyrotropin stimulates iodide uptake but not sodium/iodide symporter messenger RNA and protein expression

Follicle-forming cat thyroid cell lines synthesizing extracellular matrix and basal membrane components: a new tool for the study of thyroidal morphogenesis

Inhibition of Iodine Organification and Regulation of Follicular Size in Rat Thyroid Tissue In Vitro

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