Following cell expansion, 1 × 105 CD29+ cells were seeded in a 96-well U-bottom plate (IWAKI, Chiyoda City, Japan) to form spheroids. Subsequently, the plate was centrifuged at 4 × 400× g for 3 min. After 3 days of incubation, spheroids were cultured in a myogenic differentiation medium comprising DMEM with GlutaMax (Life Technologies) supplemented with 5% horse serum (Life Technologies) for 5 days. Next, spheroids were cultured in adipogenic differentiation medium comprising DMEM with GlutaMax (Life Technologies), supplemented with 5% horse serum (Life Technologies), 1 μM IBMX, 1 μM dexamethasone, 1 mM insulin-transferrin-selenium, 50 μM indomethacin, and 500 μM oleic acid (Tokyo Chemical Industry) for 3 days. Subsequently, the medium was replaced with DMEM supplemented with GlutaMax (Life Technologies), 5% horse serum (Life Technologies), 1 μM IBMX, 1 μM dexamethasone, 50 μM indomethacin, and 500 μM oleic acid. Cells were cultured for 2 weeks to facilitate adipogenic differentiation. All chemicals used in the experiment were for research purposes only.
Indomethacin
Manufactured by Tokyo Chemical Industry
Sourced in Japan
Indomethacin is a non-steroidal anti-inflammatory drug (NSAID) that is commonly used in pharmaceutical research and development. It is a white, crystalline powder with a melting point of approximately 160°C. Indomethacin is soluble in organic solvents such as ethanol, acetone, and chloroform.
Automatically generated - may contain errors
Lab products found in correlation
Kollidon va64, by BASF (2 mentions)
Horse serum, by Thermo Fisher Scientific (1 mentions)
Oleic acid, by Tokyo Chemical Industry (1 mentions)
Glutamax, by Thermo Fisher Scientific (1 mentions)
Insulin, by Thermo Fisher Scientific (1 mentions)
Pen strep, by Thermo Fisher Scientific (1 mentions)
High glucose dmem, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Dexamethasone (dex), by Merck Group (1 mentions)
Ckx53, by Olympus (1 mentions)
Hydrochloric acid (hcl), by Thermo Fisher Scientific (1 mentions)
Potassium dihydrogen phosphate, by Thermo Fisher Scientific (1 mentions)
Orthophosphoric acid, by Thermo Fisher Scientific (1 mentions)
Acetonitrile, by Thermo Fisher Scientific (1 mentions)
Dipotassium hydrogen orthophosphate, by Thermo Fisher Scientific (1 mentions)
Pva 4 88, by Merck Group (1 mentions)
Parteck mxp, by Merck Group (1 mentions)
5 aminosalicylic acid 5 asa, by Merck Group (1 mentions)
Paracetamol, by Merck Group (1 mentions)
Poly ε caprolactone pcl, by Merck Group (1 mentions)
10 protocols using indomethacin
1
Adipogenic Differentiation of CD29+ Cells
2
Adipogenic Differentiation of hASCs
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To investigate the effect of NHDC and CTP on adipogenic differentiation, differentiation of hASCs into adipocytes was induced. hASCs were seeded at a cell density of 4.0 × 105 cells/well in the proliferation medium (as previously described, MesenPRO RSTM Medium supplemented with growth factor and 2 mM of l -glutamine) for 5 days, and treated with adipogenic differentiation medium composed of DMEM-high glucose (Gibco, Paisley, Scotland), 10% fetal bovine serum (FBS; Gibco, Paisley, Scotland), 1% penicillin-streptomycin (Pen-strep; Gibco, Grand Island, NY, USA), 10 μg/mL insulin (Gibco, Paisley, Scotland), 500 μM isobutyl-methylxanthine (IBMX; Santa Cruz Biotechnology, CA, USA), 200 μM indomethacin (Tokyo Chemical Industry Co., Ltd., Tokyo, Japan), and 1 μM dexamethasone (Sigma-Aldrich, St. Louis, MO, USA) for another 14 days. 4 μM/L of NHDC and CTP were treated with differentiation medium as needed. Each proliferation and differentiation medium was changed every three days. The cell morphology was observed and photographed under an inverted fluorescence microscope (Olympus CKX53, Tokyo, Japan). hASCs were cultured in proliferation and differentiation medium in the presence or absence of 4 μM/L of NHDC or CTP.
3
Clay Mineral-Based Indomethacin Formulation
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Veegum F ® (Magnesium aluminium metasilicate) and Laponite RDS ® (Lithium magnesium sodium silicate) clay minerals were kindly donated by Vanderbilt minerals llc. (USA) and BYK additives ltd. (Germany) respectively. Indomethacin was purchased from Tokyo chemical industries (Japan), with a purity of >98.0% and all the reagents were used as received. Other chemical reagents such as hydrochloric acid, di-Potassium hydrogen orthophosphate, Potassium di-hydrogen phosphate, acetonitrile (HPLC grade), Ortho-phosphoric acid were purchased from Fisher scientific UK and used as received.
4
Indomethacin Formulation Development
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Indomethacin was produced by Tokyo Chemical Industry Co., LTD (6–15-19, Toshima, Kitaku, Tokyo, Japan) and provided by Merck KGaA (Darmstadt, Germany). Silica based effect pigment (Candurin ® NXT Ruby Red), silicon dioxide colloidal (EMPROVE® ESSENTIAL, Ph. Eur., JP, NF, E 551), P1 (Parteck® MXP, PVA 3–82), P2 (PVA 5–74), and P3 (Parteck® MXP, PVA-4-88) were kindly provided by Merck KGaA (Darmstadt, Germany). P4 (Kollidon ® VA 64, copovidone) was kindly provided by BASF SE (Ludwigshafen, Germany). P5 (Plasdone™ S-630, copovidone) was kindly provided by Ashland Industries Europe GmbH (Schaffhausen, Switzerland).
5
Fabrication of Silicone Elastomer Devices
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Sylgard® 184 two part Silicone Elastomer (viscosity (mixed)=3500 cP, specific gravity (cured)=1.03) containing the prepolymer base and the curing agent was obtained from Dow Corning (MI, USA) and Silicon (Si) wafers, 4inch (100) single-side polished, were obtained from Topsil GlobalWafers A/S (Frederikssund, Denmark). SU-8 2035, 2075 and SU-8 developer were purchased from micro resist technology GmbH (Berlin, Germany). Poly-εcaprolactone (PCL) (Mn = 80,000 g mol -1 ) was purchased from Sigma Aldrich (MO, USA), and HTM 140M V2 3D printing photopolymer from EnvisionTEC GmbH (Gladbeck, Germany). 5-Aminosalicylic acid (5-ASA) and paracetamol were supplied by Sigma Aldrich and indomethacin from Tokyo Chemical Industry (Tokyo, Japan). Phosphate buffered saline (PBS) was obtained from Sigma Aldrich.
6
Synthesis and Purification of MPC and BMA
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MPC was purchased from NOF Corp. (Tokyo, Japan), which was synthesized and purified at an industrial level and to medical grade specifications, according to a previously reported method. 30 BMA (Kanto Kagaku, Tokyo, Japan) was reagent grade and was purified by vacuum distillation. Fractions of BMA with a boiling point of 63 °C/ 24 mmHg were collected. Benzyl alcohol, n-butanol, and toluene were purchased from Kanto Kagaku and dried using 4A molecular sieves. Benzyl methacrylate (BZMA, FUJIFILM Wako Chemicals), 2,2 0bipyridyl (Bpy, FUJIFILM Wako Chemicals), copper (I) bromide (CuBr, Merck KGaA, Darmstadt, Germany), dibutyltin(IV) dilaurate (DBTL, FUJIFILM Wako Chemicals, Osaka, Japan), indomethacin (Tokyo Chemical Industry), 2-isocyanatoethyl methacrylate (IEMA, Tokyo Chemical Industry, Tokyo, Japan), PVP (Kollidon Ò K30, BASF, Ludwigshafen am Rhein, Germany), and all other reagents and solvents were commercially available extra-pure grade and used with no further purification.
7
Solubility Enhancement of Indomethacin
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Indomethacin (IND) was supplied by Tokyo Chemical Industry Co. Ltd (Tokyo, Japan), Soluplus (polyvinyl caprolactam-polyvinyl acetate-polyvinyl-glycol graft copolymer) and Kollidon VA64 (vinylpyrrolidone-vinyl acetate copolymer) were kindly donated by BASF (Ludwigshafen, Germany). HPLC solvents (acetonitrile, acetic acid) were of analytical grade and purchased from Fisher Scientific (Loughborough, UK). All chemicals were used as received.
8
Synthesis of Selenophene-based Polymeric Prodrugs
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Isophorone diisocyanate (IPDI, 99%), dibutyltin dilaurate (DBTDL, 95%), p-hydroxybenzaldehyde (p-HBD, 98%) and p-toluenesulfonic acid (p-TSA, 98%) were purchased from Shanghai Aladdin reagent Co., LTD (Shanghai, China). YmerTM N120, a commercially available α-methoxy-ω-diol poly(ethylene glycol) (Mn = 1000), was purchased from Perstorp special chemicals (Malmö, Sweden) and vacuum-dried at 80 °C for 12 h before use. 2,2-Bis(bromomethyl)propane-1,3-diol (BMP, 98%) and indomethacin (IND, >98%) were purchased from TCI chemicals (Shanghai, China). Sodium diselenide (Na2Se2) was synthesized according to the literature [39 (link)]. Other reagents of analytical grade were purchased from Tianjin Kemiou Chemical Reagent Co., Ltd. (Tianjin, China), and used as received.
9
Accelerated Oxidation Stability Evaluation of Drugs
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N-methyl pyrrolidone (NMP) was procured from Sigma Aldrich (Steinheim, Germany). Five of the drug compounds namely, droperidol (DPD), mifepristone (MIF) cimetidine (CIM), naproxen (NAP), clotrimazole (CLO) were purchased from Merck (Sigma Aldrich®, Vienna, Austria), while indomethacin (IMC), nifedipine (NIF), and diclofenac (DIC) were obtained from TCI chemicals® (Tokyo, Japan). Olanzapine (OLA) was purchased from Dr. Reddy’s laboratories (Hyderabad, India). All the selected drugs had a chemical purity above 99.90%, and their chemical structures are shown in Figure 1 . Polyvinylpyrrolidone (PVP) K-60 was procured from Merck (Sigma Aldrich®, Vienna, Austria). The weighing of solid powders was done using a laboratory analytical balance. Ultrapure water was obtained from a TKA water purification unit (Vienna, Austria). Accelerated oxidation stability was conducted using a high temperature/pressure oxidation testing device, RapidOxy® (Anton Paar GmbH®, Graz, Austria).
10
Characterization of Indomethacin-Soluplus Dispersions
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Indomethacin was purchased from TCI Chemicals, UK. Soluplus was a kind gift from BASF, Germany. The solvents methanol and ethanol used in the preparation of the solid dispersions were of analytical grade and were purchased from Fisher (UK). Phosphate buffer (pH 7.2) was the media used for IDR or pseudo IDR determination, solubility testing as well as whole dosage imaging and was prepared according to the USP 2003 using sodium hydroxide and potassium phosphate monobasic purchased from Fisher (UK) and Acros Organics (Germany) respectively.
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