RAW cell line was acquired from American Type Culture Collection (ATCC). These cells were unfrozen and were cultured at 1 × 106 cells in a TC-25 flask (Costar, ON, Canada) with RPMI 1640 medium supplemented with 10% FBS (GIBCO BRL, Grand Island, NY, USA), 100 unit/mL penicillin, 10 μg/mL streptomycin and 250 ng/mL amphotericin B (Sigma-Aldrich; Saint Louis, MO, USA).
After a 2-day incubation period (37 °C, 5% CO2), cells were collected, and viability was determined by trypan blue exclusion using a TC-20 cell counter system (Bio-Rad, Hercules, CA, USA). The cells in culture were routinely 90 to 95% live cells. The cells were used in passages 10–15, according to previous reports [19 (link)].
After a 2-day incubation period (37 °C, 5% CO2), cells were collected, and viability was determined by trypan blue exclusion using a TC-20 cell counter system (Bio-Rad, Hercules, CA, USA). The cells in culture were routinely 90 to 95% live cells. The cells were used in passages 10–15, according to previous reports [19 (link)].
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