Chinese hamster ovary (CHO-K1) cells and a murine pituitary cell line (GH3) were obtained from the American Type Culture Collection (ATCC, Manassas, VA) and maintained in culture at 37 °C, 5% CO2 per the supplier’s recommendations. CHO-K1 cells were cultured in Ham’s F12K medium supplemented with 10% fetal bovine serum (FBS, Atlanta Biologicals, Norcross, GA), 100 IU/mL penicillin, and 0.1 μg/mL streptomycin. GH3 cells were cultured in Ham’s F12K medium supplemented with 2.5% FBS, 15% horse serum (ATCC), 100 IU/mL penicillin, and 0.1 μg/mL streptomycin. The media and its components were purchased from Mediatech Cellgro (Herndon, VA). In the passage immediately preceding experiments, cells were transferred onto glass coverslips pretreated with poly-L-lysine to improve cell adhesion.
The composition of solutions utilized in different experiments is described in respective sections below. Chemicals were purchased from Sigma-Aldrich (St. Louis, MO) and Life Technologies (Grand Island, NY). The osmolality of the solutions was between 290 and 310 mOsm/kg, as measured by a freezing point microosmometer (Advanced Instruments, Inc., Norwood, MA). All experiments were performed at room temperature (22 ± 2 °C).
The composition of solutions utilized in different experiments is described in respective sections below. Chemicals were purchased from Sigma-Aldrich (St. Louis, MO) and Life Technologies (Grand Island, NY). The osmolality of the solutions was between 290 and 310 mOsm/kg, as measured by a freezing point microosmometer (Advanced Instruments, Inc., Norwood, MA). All experiments were performed at room temperature (22 ± 2 °C).
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