Triiodothyronine

Manufactured by Merck Group

Sourced in United States, Germany, United Kingdom, France, Macao, China, Italy

Triiodothyronine is a laboratory instrument used to measure the levels of the thyroid hormone triiodothyronine in biological samples. It is a key component in the assessment of thyroid function and disorders.

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Triiodothyronine (T3) (68 citations) 3,5,3’-Triiodothyronine (T3) (8 citations) 3,3,5- triiodothyronine (6 citations) Dexamethasone (D4902), insulin (I5500), 3-IsoButyl-1-MethylXanthine (IBMX, I7018), biotin (B4639), D-pantothenic acid hemicalcium salt (P5155), isoproterenol (I6504), CL-316,243 (C5976), norepinephrine (A7257), dibutyryl-cAMP (D0260), triiodothyronine (T5516), acetylcholine chloride (A2661), nicotine (N3876), (2 citations)

174 protocols using triiodothyronine

Adipogenic Differentiation of hAPCs

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Abdominal subcutaneous adipose tissue was excised from healthy female patients undergoing abdominoplastic surgery between June 2017 and May 2018. All subjects provided written informed consent. This study was approved by the Ethics Committee of the Medical University of Vienna and was conducted in accordance with the principles of the Declaration of Helsinki (EK 1149/2011, EK 1032/2013). hAPCs were isolated and cultured as previously described 11 (link). For each experiment cells from at least three donors were isolated. For every experiment the sample size was determined empirically based on preliminary experiments. Details in Supplementary Methods.
Confluent cells were induced using differentiation media supplemented with 0,85µM insulin, 2nM triiodothyronine, 5µM rosiglitazone, 0,5mM isobutylmethylxanthine, 1µM dexamethasone (all Sigma, St. Louis, MO, USA) for 2 days, followed by post-differentiation media supplemented with 0,85µM insulin, 2nM triiodothyronine, 5µM rosiglitazone (all Sigma). The post-differentiation media was changed every other day until day 6. Cells were stimulated with human recombinant parathyroid hormone (MyBioSource, San Diego, CA, USA), isoproterenol (Sigma) or vehicle on day 6 or during the entire adipogenic differentiation.

Hedesan O.C., Fenzl A., Digruber A., Spirk K., Baumgartner-Parzer S., Bilban M., Kenner L., Vierhapper M., Elbe-Bürger A, & Kiefer F.W. (2018). Parathyroid hormone induces a browning program in human white adipocytes. International journal of obesity (2005).

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Adipogenic Differentiation of hAPCs

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Adipocyte Precursor Cell Isolation and Differentiation

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Human adipocyte precursor cells (hAPCs) were isolated from fat specimens obtained during abdominoplasty from three different donors. Abdominal subcutaneous adipose tissue was excised from healthy female patients undergoing abdominoplastic surgery between June 2017 and May 2018. All subjects provided written informed consent. This study was approved by the Ethics Committee of the Medical University of Vienna and was conducted in accordance with the principles of the Declaration of Helsinki (EK 1149/2011 and EK 1032/2013). hAPCs were isolated and cultured as previously described.⁴⁶ Confluent cells were induced (day 0) using a differentiation medium supplemented with 0.85 μM insulin (#I9278, Sigma‐Aldrich), 2 nM triiodothyronine (#I9278, Sigma‐Aldrich), 5 μM rosiglitazone (#R2408, Sigma‐Aldrich), 0.5 mM isobutylmethylxanthine (#I7018, Sigma‐Aldrich), and 1 μM dexamethasone (#D8893, Sigma‐Aldrich) for 2 days, followed by postdifferentiation medium supplemented with 0.85 μM insulin, 2 nM triiodothyronine, and 5 μM rosiglitazone. The postdifferentiation medium was changed every other day until day 6. Cells were stimulated with CL316,243 or vehicle (H₂O) during the entire adipogenic differentiation.

Philippe C., Klebermass E., Balber T., Kulterer O.C., Zeilinger M., Egger G., Dumanic M., Herz C.T., Kiefer F.W., Scheuba C., Scherer T., Fürnsinn C., Vraka C., Pallitsch K., Spreitzer H., Wadsak W., Viernstein H., Hacker M, & Mitterhauser M. (2021). Discovery of melanin‐concentrating hormone receptor 1 in brown adipose tissue. Annals of the New York Academy of Sciences, 1494(1), 70-86.

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Differentiation of Immortalized Brown Preadipocytes

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Immortalized brown preadipocyte cells (iBAs) from mouse brown adipose tissue^{25 (link)} were obtained from Dr. Bruce Spiegelman. iBAs were cultured in Dulbecco’s Modified Eagle’s medium (DMEM; Gibco, Grand Island, NY), 20% fetal bovine serum (FBS; Gibco), 1% penicillin/streptomycin solution (P/S; Gibco), and 2 μM HEPES (Gibco). Confluent iBAs were induced to differentiate from preadipocytes by adding induction cocktail containing 5 μM dexamethasone (Sigma-Aldrich, St. Louis, MO), 125 μM indomethacin (Sigma-Aldrich), 0.02 μM insulin (Sigma-Aldrich), 500 μM isobutylmethylxanthine (Life Technologies, Carlsbad, CA), 1 nM triiodothyronine (Sigma-Aldrich), and 1 μM rosiglitazone (Sigma-Aldrich) in medium consisting of DMEM, 10% FBS, and 1% P/S. After 48 h of induction, the medium was replaced by maintenance medium consisting of DMEM, 10% FBS, 1% P/S with 0.02 μM insulin, 1 nM triiodothyronine, and 1 μM rosiglitazone. The maintenance medium was replaced every 2 days. For immunostaining and confocal applications, iBAs were plated on glass coverslips, which were pre-treated with 0.01% collagen type I (Sigma-Aldrich) in 0.01 M acetic acid.

Li Y., Imai N., Nicholls H.T., Roberts B.R., Goyal S., Krisko T.I., Ang L.H., Tillman M.C., Roberts A.M., Baqai M., Ortlund E.A., Cohen D.E, & Hagen S.J. (2021). Thioesterase superfamily member 1 undergoes stimulus-coupled conformational reorganization to regulate metabolism in mice. Nature Communications, 12, 3493.

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Adipocyte Differentiation from Adipose-Derived SVCs

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Human SVCs derived from adipose tissue were cultured in DMEM/F-12 medium and grown to confluence. Adipogenic differentiation was induced by addition of a hormone cocktail in media for two days [10% fetal calf serum; isobutylmethylxanthine (Sigma), 0.5 mM; dexamethasone (Sigma), 5 µM; rosiglitazone (Cayman), 1 µM; insulin (Sigma), 5 µM; triiodothyronine (Sigma), 1 nM; indomethacin (Sigma), 125 µM] followed by maintenance media until time of harvest [DMEM/F-12 medium in the presence of rosiglitazone (Cayman), 1 µM; insulin (Sigma), 5 µM; triiodothyronine (Sigma)]. For norepinephrine treatment (NW-NE and OW-NE groups), cells were treated with 1 μM of norepinephrine during adipogenic induction, and were then collected at days 0, 2, 5, 8, 11, and 14 during adipocyte differentiation for RT-PCR analysis.

Qiu Y., Sun L., Hu X., Zhao X., Shi H., Liu Z, & Yin X. (2020). Compromised browning plasticity of primary subcutaneous adipocytes derived from overweight Chinese adults. Diabetology & Metabolic Syndrome, 12, 91.

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Retroviral Transduction of Primary Aortic SMCs

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Retrovirus production and infection from pMSCV-PRDM16 or pMSCV-GFP was performed as described previously (Seale et al., 2008 (link)). To generate stable cells, primary murine aortic SMCs were infected with the indicated retroviruses and selected with 3 µg/ml puromycin. Adipogenic differentiation was induced by addition of a hormone cocktail in media for two days [isobutylmethylxanthine (Sigma), 0.5 mM; dexamethasone (Sigma), 5 µM; rosiglitazone (Cayman), 1 µM; insulin (Sigma), 5 µM; triiodothyronine (Sigma), 1 nM; indomethacin (Sigma), 125 µM] followed by maintenance media until time of harvest [rosiglitazone (Cayman), 1 µM; insulin (Sigma), 5 µM; triiodothyronine (Sigma)]. Where indicated, cells were treated with forskolin (10 µM, Sigma) for 4 h. A detailed protocol for the isolation of primary aortic SMCs can be found in the Supplemental Experimental Procedures.

Long J.Z., Svensson K.J., Tsai L., Zeng X., Roh H.C., Kong X., Rao R.R., Lou J., Lokurkar I., Baur W., Castellot JJ J.r., Rosen E.D, & Spiegelman B.M. (2014). Ribosomal Profiling Provides Evidence for a Smooth Muscle-Like Origin of Beige Adipocytes. Cell metabolism, 19(5), 810-820.

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Culturing Human Skin Epithelial Cells

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Human skin epithelial cells (HSEC) (CRL-4048) were purchased from the American Type Culture Collection (ATCC) (Manassas, VA, USA). Cells were cultured in epithelial cell culture medium consisting of a mixture of HAM-F12 (150 mL), DMEM (300 mL) and FBS (50 mL) supplemented with penicillin/streptomycin (50 IU/mL), adenine (24 µg/mL), insulin (5 µg/mL), triiodothyronine (1.3 ng/mL), hydrocortisone (0.4 µg/mL) and epidermal growth factor (EGF) (10 mg/mL) (all from Sigma-Aldrich/Merck). The HSEC were grown at 37 °C in a humidified incubator with 5% CO₂, and the culture medium was replaced every 2–3 days.

Cases-Perera O., Blanco-Elices C., Chato-Astrain J., Miranda-Fernández C., Campos F., Crespo P.V., Sánchez-Montesinos I., Alaminos M., Martín-Piedra M.A, & Garzón I. (2022). Development of secretome-based strategies to improve cell culture protocols in tissue engineering. Scientific Reports, 12, 10003.

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Adipocyte Differentiation from Subcutaneous AT

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ASC from subcutaneous AT of obese donors were seeded at a density of 1 × 10⁵ cells per well in a 12-well plate (3.8 cm²/well) containing basal culture medium (DMEM:F-12 medium, 1:1 vol/vol supplemented with 10% FCS) for 2 d. Cells were stimulated to differentiate using an adipogenic culture medium composed of basal culture medium supplemented with 1.8 µM insulin, 0.5 mM isobutylmethylxantine (IBMX), 500 nM dexamethasone, 1 μM rosiglitazone, 2 nM triiodothyronine and 10 μg/ml transferrin (all from Sigma-Aldrich). During differentiation, the culture medium was replaced every 2 to 3 d. Differentiated adipocytes were obtained after 8–14 d. Differentiation was validated by light microscopy, Oil Red O visualization of lipid droplets.

Chehimi M., Robert M., Bechwaty M.E., Vial G., Rieusset J., Vidal H., Pirola L, & Eljaafari A. (2016). Adipocytes, like their progenitors, contribute to inflammation of adipose tissues through promotion of Th-17 cells and activation of monocytes, in obese subjects. Adipocyte, 5(3), 275-282.

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Analytical Standards for Bioactive Compounds

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The reference standard of PA (molecular weight: 464.6 g·mol⁻¹; purity: HPLC ≥ 95%, #16,386) was purchased from Cayman Chemical, Ann Arbor, MI, USA. The reference standards of 20E (molecular weight 480.64 g·mol⁻¹; purity: HPLC ≥ 95%, #89,651) and TU (molecular weight: 496.6 g·mol⁻¹; purity: HPLC ≥ 95%, #85,781) were obtained from PhytoLab GmbH & Co. KG, Vestenbergsgreuth, Germany. Analytical grade dimethyl sulfoxide (DMSO), isopropanol, acetonitrile, methanol, cell culture medium Dulbecco’s modified Eagle’s medium/Nutrient F-12 Ham, Oil red O (ORO; 0.5% solution in isopropanol), fetal bovine serum, penicillin/streptomycin 10,000 IU/10 mg·mL⁻¹, d-biotin (purity > 99%), d-pantothenic acid (purity > 99%), human apo-transferrin (purity > 98%), rosiglitazone (purity: HPLC > 98%), human insulin, 3-isobutyl-1-methylxantine (purity: HPLC > 99%), dexamethasone (purity: HPLC > 98%), triiodothyronine (purity > 95%), cortisol (purity > 95%), and isoproterenol hydrochloride (purity: HPLC > 98%) were obtained from Merck KGaA (Darmstadt, Germany).

Todorova V., Savova M.S., Ivanova S., Ivanov K, & Georgiev M.I. (2023). Anti-Adipogenic Activity of Rhaponticum carthamoides and Its Secondary Metabolites. Nutrients, 15(13), 3061.

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Adipocyte Differentiation Induction Protocol

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White and brown adipocyte differentiation was induced as described previously, with minor modifications^{26 (link)}. For white adipocyte differentiation, cells were treated in DMEM containing 10% FBS, 10 μg/ml insulin (Wako), 2.5 μM dexamethasone (Sigma Aldrich), and 500 μM 3-isobutyl-1-methylxanthine (Sigma Aldrich). After 3 days, the medium was changed to DMEM containing 10% FBS and 10 μg/ml insulin for a further 5 days. For brown adipocyte differentiation, cells were treated in DMEM containing 10% FBS, 5 μg/ml insulin, 1 μM dexamethasone, 500 μM 3-isobutyl-1-methylxanthine, 125 nM indomethacin (Wako), 1 nM triiodothyronine (Merck), and 1 μM rosiglitazone (Wako). After 3 days, the medium was changed to DMEM containing 10% FBS, 5 μg/ml insulin, 1 nM triiodothyronine, and 1 μM rosiglitazone for a further 5 days. The medium was changed every day. After fixation by 4% paraformaldehyde/PBS, the cells were stained with Oil Red O and images were evaluated by microscopy (BZ-9000; Keyence).

Ito N., Kii I., Shimizu N., Tanaka H, & Takeda S. (2017). Direct reprogramming of fibroblasts into skeletal muscle progenitor cells by transcription factors enriched in undifferentiated subpopulation of satellite cells. Scientific Reports, 7, 8097.

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