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Cells were differentiated by treating with DMEM medium containing 500 [micro]M IBMX, 1 [micro]M dexamethasone, 1 [micro]g/ml insulin, and 10% FBS for 2 days, followed by incubation with DMEM medium containing 1 [micro]g/ml insulin and 10% FBS till lipid droplets were formed.
Differentiation was induced by incubating 2-day postconfluent MSCs in an MDI induction medium (DMEM supplemented with 10% fetal bovine serum, 0.5 mM IBMX, 1 [micro]m dexamethasone, and 1 [micro]g/mL insulin) for 2 days.
When cells reached confluence (day 0), they were treated with 100 nM insulin (I) and 500 [micro]M IBMX (M) until day 4, with a media change on day 2.
Mouse oocytes were placed in drops of culture media containing 200 [micro]M IBMX for microinjection.
Numerous pigmentation enhancing agents have been discovered, such as diacylglycerols, 3-isobutyl-1-methylxanthine (IBMX), and dimethylsulfoxide (DMSO).
Chrenek, "Effect of oxytocin, IBMX and dbcAMP on Rabbit ovarian follicles," Anatomia Histologia Embryologia, vol.
After this, growth medium was substituted by differentiation medium, which consisted of the same medium supplemented with 0.5 mM 3-isobutyril-l-methylxanthine (IBMX), 1 [micro]M dexamethasone, 10 [micro]g/ml insulin and 1 [micro]M rosiglitazone.
At the 3[sup]rd day after confluence, adipocytes differentiation was induced by adding 3-Isobutyl-1-Methylxanthine (IBMX; 0.5 mmol/L), dexamethasone (0.25 [micro]mol/L), and insulin (10 mg/L) for 2 days.
Dulbecco's modified Eagle's medium (DMEM), Streptozotocin (STZ), Methanol, fetal calf serum, glucose, glutamine, penicillin, streptomycin, diazoxide, 3-Isobutyl-1-methylxanthine (IBMX), calcium chloride, potassium chloride, ethylene glycol tetra acetic acid (EGTA) and 3-(4, 5-Dimethylthiazol-2-yl)-2, 5- Diphenyltetrazolium Bromide (MTT) reagents were procured from Sigma Aldrich (Bangalore, India).
When the cultures reached confluency (day 0), cells were treated with differentiation medium containing DMEM supplemented with 10% fetal bovine serum (FBS ATCC, cat: 30-2020), 0.25 mM dexamethasone (Sigma-Aldrich -D4902), 5 mg/mL insulin (Sigma-Aldrich-I3536) and 0.5 mM isobutylmethylxanthine (IBMX, Sigma-Aldrich-I7018).
During acquisition, 50 [micro]M Forskolin (adenylate cyclase activator) is added at 5 minutes of time and 10 [micro]M Rolipram+ 500 [micro]M IBMX (phosphodiesterase inhibitors) were added at time point of 15 minutes.
Dimethyl sulfoxide and 3-isobutyl-1-methylxanthine (IBMX) were purchased from Fluka Chemical Co.