Human pulmonary epithelial A549 cells were maintained in Roswell Park Memorial Institute medium (RPMI) containing 10% FBS and penicillin/streptomycin. Cells were grown at 37°C in a humidified atmosphere of 95% air/5% CO2 and fed every 2–3 d. Before treatment, the cells were washed with phosphate-buffered saline and cultured in RPMI/5% charcoal–dextran stripped FBS (CD-FBS) for 2 d. For the hypoxic condition, cells were incubated at a CO2 level of 5% with 1% O2 balanced with N2 using a hypoxic chamber (Thermo Fisher Scientific, Waltham, MA, USA). KRG stock was prepared at 10 mg/mL in phosphate buffer saline and diluted with media to 1 mg/mL just prior to use and sterilized by filtration with a 0.22 μm bottle top filter (Thermo Fisher Scientific).
0.22 μm bottle top filter
Manufactured by Thermo Fisher Scientific
Sourced in United States
The 0.22 μm bottle top filter is a laboratory filtration device designed to remove particulates and microorganisms from liquids. It features a 0.22 μm membrane that effectively filters out particles and microbes, producing a clarified filtrate. This filter can be used to sterilize a variety of aqueous solutions.
Automatically generated - may contain errors
2 protocols using 0.22 μm bottle top filter
1
Culturing and Treating A549 Cells
2
Gelatin and Alginate Hydrogel Encapsulation of Primary Rat Schwann Cells
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Gelatin (from bovine skin), medium viscosity sodium alginate (sodium salt from brown algae, medium viscosity), calcium chloride (CaCl2), Dulbecco's modified Eagle's medium (DMEM), fetal bovine serum (FBS), trypsin, ethylene-diaminetetraacetic acid (EDTA), and fluorescent dyes (propidium iodide (PI), calcein, Hoechst 33342) were all obtained from Sigma Aldrich (St. Louis, MO). Phosphate buffered saline (PBS; 0.0067 M) was obtained from HyClone (Logan, Utah). Primary rat Schwann cells (PRSCs) were supplied by Saskatoon City Hospital. A Sartorius scale (model 225d; Shanghai, China) was used for weighing samples. A 0.22-μm bottle-top filter (Thermo Scientific, Ann Arbor, MI, USA) was used to filter alginate solutions. A freeze-dryer (FreeZone, Labconco, USA) was used to freeze-dry samples. Sterile circular coverslips (Thermo Scientific™) and 12/24-well tissue culture plates (Thermo Fisher Scientific, MA, USA) were used for experiments.
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