DS-1 cells (human corpus cavernosum cell line) were routinely cultured in a humidified 5% CO2–95% air incubator in Dulbecco's modified eagles medium (DMEM) supplemented with 10% foetal bovine serum (FBS), 20 μM l-glutamine, 1 mM pyruvate, 200 U/ml penicillin G, 200 μg/ml streptomycin and 4.5 mg/ml of d-glucose at 37 °C. Cells were allowed to attach to the bottom of a 96-well plate overnight and were then exposed to the extract (10 mg/ml PBS) or placebo PBS (control) for 24 h. The cell media was assayed for the concentrations of nitrate (NO3−) and nitrite (NO2−), the final in vivo products of NO, using a commercial nitrate reductase preparation and Griess reagent kit (Cayman Chemical, USA) with the resultant absorbance being read at 550 nm using a VictorX™ plate reader (Perkin Elmer USA)
DS-1 cells (human corpus cavernosum cell line) were routinely cultured in a humidified 5% CO2–95% air incubator in Dulbecco's modified eagles medium (DMEM) supplemented with 10% foetal bovine serum (FBS), 20 μM l-glutamine, 1 mM pyruvate, 200 U/ml penicillin G, 200 μg/ml streptomycin and 4.5 mg/ml of d-glucose at 37 °C. Cells were allowed to attach to the bottom of a 96-well plate overnight and were then exposed to the extract (10 mg/ml PBS) or placebo PBS (control) for 24 h. The cell media was assayed for the concentrations of nitrate (NO3−) and nitrite (NO2−), the final in vivo products of NO, using a commercial nitrate reductase preparation and Griess reagent kit (Cayman Chemical, USA) with the resultant absorbance being read at 550 nm using a VictorX™ plate reader (Perkin Elmer USA)
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