Stable l glutamine

Vero cells (clone E6, CCLV-RIE-929/25) were cultivated in MEM with Earle’s salts (Capricorn Scientific, Ebsdorfergrund, Germany) supplemented with 2 mM stable L-glutamine (Capricorn Scientific), 100 U/mL penicillin, 100 µg/mL streptomycin (Capricorn Scientific), and 10% fetal bovine serum (FBS) (Capricorn Scientific) at 37 °C in 5% CO₂. C6/36 (CCLV-RIE-1299) cells were grown in Schneiders Drosophila media (PAN-Biotech, Aidenbach, Germany) supplemented with 2 mM stable L-glutamine (Capricorn Scientific), 1 mM sodium pyruvate (PAN Biotech), 1X MEM NEAA (PAN Biotech), 100 U/mL penicillin, 100 µg/mL streptomycin (Capricorn Scientific), and 10% FBS (Biowest, Riverside, MO, USA) at 28 °C. The suspension cell line FreeStyle™ CHO-S (Thermo Fisher Scientific, Waltham, MA, USA) was cultivated in FreeStyle^TM CHO medium (Thermo Fisher Scientific) supplemented with 8 mM stable L-glutamine (Capricorn Scientific), 100 U/mL penicillin, 100 µg/mL streptomycin (Capricorn Scientific) at 37 °C, and 5% CO₂ on an orbital shaker.

MDA-MB-231 and MCF-7 cells were grown as adherent monolayers in low glucose (1.0 g/L) or high glucose (5 g/L) Dulbecco's Modified Eagle's Medium (DMEM) with stable L-glutamine (2 mM) and sodium pyruvate (from CAPRICORN scientific) supplemented with 10% heat-inactivated fetal bovine serum, 10 μg/mL human recombinant Zn-insulin, and antibiotics: penicillin (100 U/ml), streptomycin (100 μg/ml) and gentamicin at a final concentration of 50 μg/ml. Cells were grown at 37°C in humidified incubator containing 5% CO₂ in air and were sub-cultured at 90% confluence.