The study was performed on Rat Pancreatic Acinar Tumor Cell Line—AR42J cells (American Type Culture Collection, Rockville, MD, USA). Cell culture was grown in the RPMI 1640 Medium supplemented with Glutamax-I (Gibco BRL, Gaithersburg, MD, USA) and 10% fetal bovine serum (FBS, heat-inactivated; Gibco-BRL, Grand Island, NY, USA) with addition of 100 U/mL penicillin and 100 µg/mL streptomycine (Sigma-Aldrich, St. Louis, MO, USA) in the standard conditions: 37 °C and 5% CO2 [127 (link),128 (link),129 (link),130 (link)].
Twenty-four hours before the experiments, culture medium was replenished with fresh RPMI 1640 containing 2% FBS and without antibiotics. Cells cultures were plated at the initial density of 2 × 106/mL in a 100 mm culture plate (Falcon 3047; Becton Dickinson, Lincoln Park, NJ, USA) and allowed to attach for 12 h. AR42J cells were incubated in GHRL-free or GHRL containing medium and in the presence or absence of caerulein or in a combination of the above. GHRL was given at a concentration of 10−7 M, caerulein at a concentration of 10−8 M, incubation time was 48 h. All experiments were repeated at least six times.
Twenty-four hours before the experiments, culture medium was replenished with fresh RPMI 1640 containing 2% FBS and without antibiotics. Cells cultures were plated at the initial density of 2 × 106/mL in a 100 mm culture plate (Falcon 3047; Becton Dickinson, Lincoln Park, NJ, USA) and allowed to attach for 12 h. AR42J cells were incubated in GHRL-free or GHRL containing medium and in the presence or absence of caerulein or in a combination of the above. GHRL was given at a concentration of 10−7 M, caerulein at a concentration of 10−8 M, incubation time was 48 h. All experiments were repeated at least six times.
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