At 8 or 18 days post-differentiation, hiPSC-CMs were dissociated with 0.025% trypsin (PromoCell Detach 2 kit), plated on matrigel-coated coverslips, and cultured in basal media for 48 h. Ten or twenty days post-differentiation, the CMs were loaded with a calcium-sensitive fluorescent dye (Fura-2 AM, cell permeant; ThermoFisher, Rockville, MD) and the ratios of fluorescence intensities (excitation ratio of 340/380 nm) were recorded using the IonOptix system (Ionoptix, Milton, MA). The electrically-induced calcium transients were triggered by pulses from a MyoPacer (IonOptix, Milton, MA) at 40 V and 0.5 Hz and measurements were obtained at room temperature. Calcium traces were analyzed using IonWizard software (IonOptix) to calculate the baseline, amplitude (peak height), and tau (time of relaxation). For measurements with isoproterenol (DL isoproterenol hydrochloride; Sigma-Aldrich, St. Louis, MO), basal calcium transients were recorded, isoproterenol (final concentration of 500 nM) was added to the cells, and then calcium transients were measured again.
Dl isoproterenol hydrochloride
Manufactured by Merck Group
Sourced in United States
DL isoproterenol hydrochloride is a synthetic catecholamine compound. It functions as a nonselective beta-adrenergic agonist, stimulating both beta-1 and beta-2 adrenergic receptors.
Automatically generated - may contain errors
Lab products found in correlation
Fura 2 am, by Thermo Fisher Scientific (4 mentions)
Ionwizard software, by IonOptix (2 mentions)
Detach 2 kit, by PromoCell (2 mentions)
Myopacer, by IonOptix (2 mentions)
Sodium hydroxide (naoh), by Merck Group (1 mentions)
Sodium chloride (nacl), by Merck Group (1 mentions)
Phosphoric acid, by Merck Group (1 mentions)
Sodium carbonate, by Merck Group (1 mentions)
Copper sulphate, by Merck Group (1 mentions)
Disodium hydrogen orthophosphate, by Merck Group (1 mentions)
Potassium dihydrogen orthophosphate, by Merck Group (1 mentions)
Phenol reagent, by Merck Group (1 mentions)
Alzet model 1002, by Durect (1 mentions)
6 protocols using dl isoproterenol hydrochloride
1
Calcium Dynamics in hiPSC-Derived Cardiomyocytes
2
Comprehensive Biochemical Assay Protocol
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B&K Technology Group China Co., Ltd. provided a standard SAC sample (Xiamen, China). The CK-MB and LDH kits were provided by Crest Biosystems and Coral Clinical Systems in Goa, India. The DL-isoproterenol hydrochloride was given by Sigma Aldrich in the United States. Nice Chemicals Pvt Ltd. in Cochin, India, provided the EDTA, and Hong Yang Chemical Corp. in China provided the ethanol. Heparin was provided by Gland Pharmaceutical Ltd. in Hyderabad, India. Hydrogen peroxide, hydroxylamine HCL, and ketamine were provided by Prem Pharmaceutical Ltd. Indore, India. Malondialdehyde (MDA), n-Butanol, and nitro blue tetrazolium were supplied by Loba Chemicals in Mumbai, India (NBT). Standard bovine albumin and sucrose were provided by S D Fine Chemicals in Mumbai, India. The thiobarbituric acid and xylazine were provided by Indian Immunological in Guntur, India. Copper sulphate, disodium hydrogen orthophosphate, phenol reagent, phosphoric acid, potassium dihydrogen orthophosphate, sodium chloride, sodium hydroxide, and sodium carbonate were given by Merck Specialties Private Limited in Mumbai, India. All of the other chemicals used in this experiment were obtained from well-known suppliers. Qualigens in Mumbai, India, and INCO in India sold AutoAnalyzer. Before being used, all equipment and instruments used in this study were calibrated.
3
Osmotic Mini-Pump Implantation for Isoproterenol Delivery in Mice
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After an acclimatisation period of 7 days, mice were subjected to subcutaneous implantation of osmotic mini-pumps (ALZET model 1002, Durect Corporation, Cupertino, California, USA; supplied by Charles River UK Ltd., Kent, UK). After an incision was made on the back, slightly posterior to the scapulae, a distal subcutaneous pocket was created and the pump was located posterior to the right flank. ISO (DL-Isoproterenol hydrochloride, Sigma, St. Louis, MO, USA) was dissolved in 0.9% NaCl at a concentration calculated to deliver 30 μg/g/day for up to 15 days at an infusion rate of 0.25 μl/h. Pumps were filled with either the ISO solution or 0.9% NaCl (vehicle) and activated according to the manufacturer's instructions. During the surgical intervention, mice were anesthetised with 3% isoflurane mixed with 97% O2 (flow rate 1 L/min) and the mini-pumps were implanted via a 0.5 cm interscapular incision under sterile surgical conditions. The wound was closed with an interrupted suture using a 6–0 silk thread.
4
Calcium Dynamics in hiPSC-Derived Cardiomyocytes
5
Isoproterenol-induced cardiac hypertrophy
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WT and HOM mice received a continuous infusion of ISO (30 mg/kg/day) or vehicle (0.9% NaCl) for 14 days via subcutaneously-implanted osmotic minipumps (ALZET model 1002; supplied by Charles River UK Ltd, Kent, UK), as described previously.21 (link) Echocardiographic recordings were obtained prior to minipump implantation and at the end of the infusion period, and hearts were collected for gravimetric, histological and biochemical analyses. ISO (DL-Isoproterenol hydrochloride) was from Sigma (MO, USA).
6
Garlic Oil's Cardioprotective Effects
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Sigma Aldrich (USA) provided the garlic oil, which was GC-MS standardized for the presence of DADS. All of the compounds utilized in this investigation were analytical grade and came from a standard source. The CK-MB and LDH kits were provided by Crest Biosystems and Coral Clinical Systems (Goa, India). The DL-isoproterenol hydrochloride was provided by Sigma Aldrich (St. Louis, MO, USA). All chemicals for the project were purchased from Al-Majharia international trading establishment in Saudi Arabia, while glassware and other required equipment were provided by Gulf Scientific Glass Industry limited in Saudi Arabia and Scientific equipment trading Establishment in Saudi Arabia, respectively.
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