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Endocrinology, doi:10.1210/endo-105-1-80
Endocrinology Vol. 105, No. 1 80-85
Copyright © 1979 by the Endocrine Society.
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Depletion of L-3,5,3'-Triiodothyronine and L-Thyroxine in Euthyroid Calf Serum for Use in Cell Culture Studies of the Action of Thyroid Hormone*

HERBERT H. SAMUELS, FREDERICK STANLEY and JUAN CASANOVA

Endocrine Division, Department of Medicine, New York University Medical Center New York, New York 10016

Address requests for reprints to: Dr. Herbert H. Samuels, Department of Medicine, Attention: C and D Building, New York University Medical Center, 550 First Avenue, New York, New York 10016.

Abstract

GH1 cells are a clonal strain of rat pituitary tumor cells which synthesize GH and PRL. We have previously demonstrated that these cells respond to physiological concentrations of L-T3 and L-T4 when cultured with medium supplemented with thyroidectomized calf serum to achieve a thyroid hormonedepleted state under cell culture conditions. In this study, we describe a method to deplete euthyroid calf serum of L-T3 and L-T4 using an anion exchange resin. We demonstrate that the procedure only minimally alters the low molecular weight anion components of the serum and does not change the total protein content or the electrophoretic pattern of serum proteins. Moreover, we show that euthyroid calf serum depleted of L-T4 and L-T4 by this procedure yields serum which, when used as a medium supplement, results in biological responses identical to those obtained with media supplemented with thyroidectomized calf serum. In addition, resin treatment does not alter the growth-promoting properties of the serum if the thyroid hormone concentration is restored. This procedure should be useful in preparing thyroid hormone-depleted serum for cell culture studies in situations where thyroidectomy is not feasible or would require surgical procedures on a large number of small animals.

Footnotes

* This work was supported by NIH Grant AM-16636 and Biomedical Research Support Grant BR-05399.

Received November 15, 1978.




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Thyrotropin Receptors in Brown Adipose Tissue: Thyrotropin Stimulates Type II Iodothyronine Deiodinase and Uncoupling Protein-1 in Brown Adipocytes
Endocrinology, March 1, 2001; 142(3): 1195 - 1201.
[Abstract] [Full Text] [PDF]

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 EndocrinologyHome page
W.-X. Gu, P. H. Stern, L. D. Madison, and G.-G. Du
Mutual Up-Regulation of Thyroid Hormone and Parathyroid Hormone Receptors in Rat Osteoblastic Osteosarcoma 17/2.8 Cells
Endocrinology, January 1, 2001; 142(1): 157 - 164.
[Abstract] [Full Text] [PDF]

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 Mol. Cell. Biol.Home page
G. R. Williams
Cloning and Characterization of Two Novel Thyroid Hormone Receptor beta Isoforms
Mol. Cell. Biol., November 15, 2000; 20(22): 8329 - 8342.
[Abstract] [Full Text]

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 EndocrinologyHome page
H. Robson, T. Siebler, D. A. Stevens, S. M. Shalet, and G. R. Williams
Thyroid Hormone Acts Directly on Growth Plate Chondrocytes to Promote Hypertrophic Differentiation and Inhibit Clonal Expansion and Cell Proliferation
Endocrinology, October 1, 2000; 141(10): 3887 - 3897.
[Abstract] [Full Text] [PDF]

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 EndocrinologyHome page
K.-h. Lin, H.-y. Shieh, and H.-C. Hsu
Negative Regulation of the Antimetastatic Gene Nm23-H1 by Thyroid Hormone Receptors
Endocrinology, July 1, 2000; 141(7): 2540 - 2547.
[Abstract] [Full Text] [PDF]

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 Cancer Res.Home page
V. Martín, M. L. Cortés, Pablo de Felipe, A. Farsetti, N. B. Calcaterra, and M. Izquierdo
Cancer Gene Therapy by Thyroid Hormone-mediated Expression of Toxin Genes
Cancer Res., June 1, 2000; 60(12): 3218 - 3224.
[Abstract] [Full Text]

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 Am. J. Physiol. Endocrinol. Metab.Home page
A. Hernandez and M. J. Obregon
Triiodothyronine amplifies the adrenergic stimulation of uncoupling protein expression in rat brown adipocytes
Am J Physiol Endocrinol Metab, May 1, 2000; 278(5): E769 - E777.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
K. Nakajima, A. Takahashi, and Y. Yaoita
Structure, Expression, and Function of the Xenopus laevis Caspase Family
J. Biol. Chem., March 31, 2000; 275(14): 10484 - 10491.
[Abstract] [Full Text] [PDF]

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 J. Nutr.Home page
M. P. Sciaudone, S. Chattopadhyay, and H. C. Freake
Chelation of Zinc Amplifies Induction of Growth Hormone mRNA Levels in Cultured Rat Pituitary Tumor Cells
J. Nutr., January 1, 2000; 130(2): 158 - 163.
[Abstract] [Full Text]

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 Mol. Cell. Biol.Home page
F. Casas, P. Rochard, A. Rodier, I. Cassar-Malek, S. Marchal-Victorion, R. J. Wiesner, G. Cabello, and C. Wrutniak
A Variant Form of the Nuclear Triiodothyronine Receptor c-ErbAalpha 1 Plays a Direct Role in Regulation of Mitochondrial RNA Synthesis
Mol. Cell. Biol., December 1, 1999; 19(12): 7913 - 7924.
[Abstract] [Full Text] [PDF]

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 Mol. Endocrinol.Home page
J. D. Furlow and D. D. Brown
In Vitro and in Vivo Analysis of the Regulation of a Transcription Factor Gene by Thyroid Hormone during Xenopus laevis Metamorphosis
Mol. Endocrinol., December 1, 1999; 13(12): 2076 - 2089.
[Abstract] [Full Text]

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 J. Immunol.Home page
M. P. Foster, E. Montecino-Rodriguez, and K. Dorshkind
Proliferation of Bone Marrow Pro-B Cells Is Dependent on Stimulation by the Pituitary/Thyroid Axis
J. Immunol., December 1, 1999; 163(11): 5883 - 5890.
[Abstract] [Full Text] [PDF]

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 EndocrinologyHome page
K. K. Jacob and F. M. Stanley
CCAAT/Enhancer-Binding Protein {alpha} Is a Physiological Regulator of Prolactin Gene Expression
Endocrinology, October 1, 1999; 140(10): 4542 - 4550.
[Abstract] [Full Text]

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 J. Biol. Chem.Home page
R. J. Denver, L. Ouellet, D. Furling, A. Kobayashi, Y. Fujii-Kuriyama, and J. Puymirat
Basic Transcription Element-binding Protein (BTEB) Is a Thyroid Hormone-regulated Gene in the Developing Central Nervous System. EVIDENCE FOR A ROLE IN NEURITE OUTGROWTH
J. Biol. Chem., August 13, 1999; 274(33): 23128 - 23134.
[Abstract] [Full Text] [PDF]

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 Mol. Cell. Biol.Home page
C. Cenciarelli, F. De Santa, P. L. Puri, E. Mattei, L. Ricci, F. Bucci, A. Felsani, and M. Caruso
Critical Role Played by Cyclin D3 in the MyoD-Mediated Arrest of Cell Cycle during Myoblast Differentiation
Mol. Cell. Biol., July 1, 1999; 19(7): 5203 - 5217.
[Abstract] [Full Text] [PDF]

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 Mol. Endocrinol.Home page
G. M. Wahlström, B. Vennström, and M. B. Bolin
The Adenovirus E1A Protein Is a Potent Coactivator for Thyroid Hormone Receptors
Mol. Endocrinol., July 1, 1999; 13(7): 1119 - 1129.
[Abstract] [Full Text]

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 Am. J. Physiol. Lung Cell. Mol. Physiol.Home page
S. Chalaka, D. H. Ingbar, R. Sharma, Z. Zhau, and C. H. Wendt
Na+-K+-ATPase gene regulation by glucocorticoids in a fetal lung epithelial cell line
Am J Physiol Lung Cell Mol Physiol, July 1, 1999; 277(1): L197 - L203.
[Abstract] [Full Text] [PDF]

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 Am. J. Physiol. Heart Circ. Physiol.Home page
D. R. Wagner, T. Kubota, V. J. Sanders, C. F. McTiernan, and A. M. Feldman
Differential regulation of cardiac expression of IL-6 and TNF-alpha by A2- and A3-adenosine receptors
Am J Physiol Heart Circ Physiol, June 1, 1999; 276(6): H2141 - H2147.
[Abstract] [Full Text] [PDF]

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 Am. J. Respir. Cell Mol. Bio.Home page
G. Otulakowski, B. Rafii, H. R. Bremner, and H. O'Brodovich
Structure and Hormone Responsiveness of the Gene Encoding the alpha -Subunit of the Rat Amiloride-Sensitive Epithelial Sodium Channel
Am. J. Respir. Cell Mol. Biol., May 1, 1999; 20(5): 1028 - 1040.
[Abstract] [Full Text]

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 EndocrinologyHome page
S. Cecconi, N. Rucci, M. L. Scaldaferri, M. P. Masciulli, G. Rossi, C. Moretti, M. D’Armiento, and S. Ulisse
Thyroid Hormone Effects on Mouse Oocyte Maturation and Granulosa Cell Aromatase Activity
Endocrinology, April 1, 1999; 140(4): 1783 - 1788.
[Abstract] [Full Text]

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 J. Biol. Chem.Home page
B. Belandia, M. J. Latasa, A. Villa, and A. Pascual
Thyroid Hormone Negatively Regulates the Transcriptional Activity of the beta -Amyloid Precursor Protein Gene
J. Biol. Chem., November 13, 1998; 273(46): 30366 - 30371.
[Abstract] [Full Text] [PDF]

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 EndocrinologyHome page
B. Yusta, E. T. Alarid, D. F. Gordon, E. C. Ridgway, and P. L. Mellon
The Thyrotropin {beta}-Subunit Gene Is Repressed by Thyroid Hormone in a Novel Thyrotrope Cell Line, Mouse T{alpha}T1 Cells
Endocrinology, November 1, 1998; 139(11): 4476 - 4482.
[Abstract] [Full Text] [PDF]

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 Proc. Natl. Acad. Sci. USAHome page
E. Carosa, A. Fanelli, S. Ulisse, R. Di Lauro, J. E. Rall, and E. A. Jannini
Ciona intestinalis nuclear receptor 1: A member of steroid/thyroid hormone receptor family
PNAS, September 15, 1998; 95(19): 11152 - 11157.
[Abstract] [Full Text] [PDF]

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 Mol. Endocrinol.Home page
P. Bartunek and M. Zenke
Retinoid X Receptor and c-erbA/Thyroid Hormone Receptor Regulate Erythroid Cell Growth and Differentiation
Mol. Endocrinol., September 1, 1998; 12(9): 1269 - 1279.
[Abstract] [Full Text]

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 J. Immunol.Home page
H.-Y. Lin, L. J. Martino, B. D. Wilcox, F. B. Davis, J. K. Gordinier, and P. J. Davis
Potentiation by Thyroid Hormone of Human IFN-{gamma}-Induced HLA-DR Expression
J. Immunol., July 15, 1998; 161(2): 843 - 849.
[Abstract] [Full Text] [PDF]

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 EndocrinologyHome page
M. J. Latasa, B. Belandia, and A. Pascual
Thyroid Hormones Regulate {beta}-Amyloid Gene Splicing and Protein Secretion in Neuroblastoma Cells
Endocrinology, June 1, 1998; 139(6): 2692 - 2698.
[Abstract] [Full Text] [PDF]

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 Mol. Endocrinol.Home page
G. M. Wahlström and B. Vennström
Requirements for Repression of Retinoid X Receptor by the Oncoprotein P75gag-v-erbA and the Thyroid Hormone Receptors
Mol. Endocrinol., May 1, 1998; 12(5): 645 - 653.
[Abstract] [Full Text]

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 EndocrinologyHome page
J. M. Staton and P. J. Leedman
Posttranscriptional Regulation of Thyrotropin {beta}-Subunit Messenger Ribonucleic Acid by Thyroid Hormone in Murine Thyrotrope Tumor Cells: A Conserved Mechanism across Species
Endocrinology, March 1, 1998; 139(3): 1093 - 1100.
[Abstract] [Full Text] [PDF]

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 EndocrinologyHome page
A. Hernandez, D. L. St. Germain, and M. J. Obregon
Transcriptional Activation of Type III Inner Ring Deiodinase by Growth Factors in Cultured Rat Brown Adipocytes
Endocrinology, February 1, 1998; 139(2): 634 - 639.
[Abstract] [Full Text] [PDF]

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 Circ. Res.Home page
D. R. Wagner, A. Combes, C. McTiernan, V. J. Sanders, B. Lemster, and A. M. Feldman
Adenosine Inhibits Lipopolysaccharide-Induced Cardiac Expression of Tumor Necrosis Factor-{alpha}
Circ. Res., January 23, 1998; 82(1): 47 - 56.
[Abstract] [Full Text] [PDF]

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 JCBHome page
Y. Su, Y. Shi, M. A. Stolow, and Y.-B. Shi
Thyroid Hormone Induces Apoptosis in Primary Cell Cultures of Tadpole Intestine: Cell Type Specificity and Effects of Extracellular Matrix
J. Cell Biol., December 15, 1997; 139(6): 1533 - 1543.
[Abstract] [Full Text] [PDF]

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 EndocrinologyHome page
R. El Meskini, F. Boudouresque, and L'H. Ouafik
Estrogen Regulation of Peptidylglycine {alpha}-Amidating Monooxygenase Messenger Ribonucleic Acid Levels by a Nuclear Posttranscriptional Event
Endocrinology, December 1, 1997; 138(12): 5256 - 5265.
[Abstract] [Full Text] [PDF]

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 Circ. Res.Home page
C. F. McTiernan, B. H. Lemster, C. Frye, S. Brooks, A. Combes, and A. M. Feldman
Interleukin-1ß Inhibits Phospholamban Gene Expression in Cultured Cardiomyocytes
Circ. Res., October 19, 1997; 81(4): 493 - 503.
[Abstract] [Full Text]

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 Proc. Natl. Acad. Sci. USAHome page
C. C. Thompson and M. C. Bottcher
The product of a thyroid hormone-responsive gene interacts with thyroid hormone receptors
PNAS, August 5, 1997; 94(16): 8527 - 8532.
[Abstract] [Full Text] [PDF]

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 Mol. Pharmacol.Home page
K. A. O'Leary, H.-C. Li, P. A. Ram, P. McQuiddy, D. J. Waxman, and C. B. Kasper
Thyroid Regulation of NADPH:Cytochrome P450 Oxidoreductase: Identification of a Thyroid-Responsive Element in the 5'-Flank of the Oxidoreductase Gene
Mol. Pharmacol., July 1, 1997; 52(1): 46 - 53.
[Abstract] [Full Text]

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 Mol. Endocrinol.Home page
F. Pernasetti, L. Caccavelli, C. Van de Weerdt, J. A. Martial, and M. Muller
Thyroid Hormone Inhibits the Human Prolactin Gene Promoter by Interfering with Activating Protein-1 and Estrogen Stimulations
Mol. Endocrinol., June 1, 1997; 11(7): 986 - 996.
[Abstract] [Full Text]

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 J. Clin. Endocrinol. Metab.Home page
M. Behr, D. B. Ramsden, and U. Loos
Deoxyribonucleic Acid Binding and Transcriptional Silencing by a Truncated c-erbA{beta}1 Thyroid Hormone Receptor Identified in a Severely Retarded Patient with Resistance to Thyroid Hormone
J. Clin. Endocrinol. Metab., April 1, 1997; 82(4): 1081 - 1087.
[Abstract] [Full Text] [PDF]

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 Mol. Endocrinol.Home page
K. Busch, B. Martin, A. Baniahmad, R. Renkawitz, and M. Muller
At Least Three Subdomains of v-erbA Are Involved in Its Silencing Function
Mol. Endocrinol., March 1, 1997; 11(3): 379 - 389.
[Abstract] [Full Text]

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 J. Biol. Chem.Home page
Y. Yaoita and K. Nakajima
Induction of Apoptosis and CPP32 Expression by Thyroid Hormone in a Myoblastic Cell Line Derived from Tadpole Tail
J. Biol. Chem., February 21, 1997; 272(8): 5122 - 5127.
[Abstract] [Full Text] [PDF]


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