Rpmi 1640 medium with l glutamine

HER2+ breast cancer cell lines BT474 (HTB-20) representing Luminal B (ER+; PR+, HER2+; ductal carcinoma) subtype and SKBR3 (HTB-30) representing HER2+ subtype (ER-; PR-; HER2+; adenocarcinoma) of breast cancer were purchased from the American Type Culture Collection (ATCC) [25 (link),26 (link)], which were known to well characterized in previous studies and decribed highly sensitive to trastuzumab treatment [27 (link),28 ]. SKBR3 cells were maintained in Mc Coy’s 5A medium (Lonza) with L-glutamine containing 10% fetal bovine serum (FBS), 1% penicillin-streptomycin. BT474 cells were maintained in RPMI 1640 medium with L-glutamine (Lonza) supplemented with 10% FBS, 1% penicillin-streptomycin and 2% bovine insulin. The cell lines were cultured in a humidified air supplemented with 5% CO₂ at 37°C. Trastuzumab was (kindly gifted by Prof. Dr. Hakan Gürdal from the Department of Pharmacology in Medical School of Ankara University) dissolved in phosphate-buffered saline (PBS) (stock concentration of 300 mg/mL) and stored at 4°C.

BHK-21 cells (ATCC-CCL10) were grown as described [31 (link)]. LLC cells were grown in DMEM (GIBCO BRL, UK) supplemented with 10% FBS, 2 mM glutamine, and antibiotics. LLC cells expressing mouse GM-CSF (LLC-GMCSF) were generated by lentivector transduction following published methodologies [9 (link), 15 –17 (link)]. MC38 cells were provided by Dr. Karl E. Hellström (University of Washington, Seattle, WA). These cells were cultured in RPMI-1640 medium (Lonza, Switzerland) supplemented with 10% FBS, 2 mM glutamine, 20 mM HEPES, antibiotics and 50 µM 2-mercaptoethanol. H1299 cells (ATCC- CRL5803) and A549 cells (ATCC-CCL-185) were purchased from the ATCC and cultured in RPMI 1640 Medium with L-Glutamine (Lonza) supplemented with 5% fetal bovine serum (FBS) and 5% Penicillin-Streptomycin 10,000 U/ml (GIBCO BRL, UK). All cell lines were tested once a month for mycoplasma by PCR.