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2 protocols using wi 38

1

Culturing Immortalized Ovarian and Colorectal Cancer Cells

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The immortalized human ovarian surface epithelial cells T29 and their transformed counterparts T29Kt1 (with the introduction of KRASV12 into the immortalized cells) were reported previously28 (link) and were cultured in medium consisting of 1:1 MCDB105 medium and M199 medium (Sigma-Aldrich Co.) with 10% fetal bovine serum (HyClone) in the presence of 1% penicillin and streptomycin (HyClone). HCT-116 (p53+/+) and HCT-116 (p53−/−) were kindly provided by Dr Chuangui Wang (East China Normal University, Shanghai, China) and were maintained in DMEM with the same supplements. The culture medium of other cells used is listed in Supplementary Table 2. A549, H441, H358, H1299, H1975, PC9, H661, H522, HCT-15, LoVo, SW480, SW620, DLD-1, HT29, CaCo2, HCT-8, CCD-18Co, CCD841CoN, Panc-1, CFPAC-1 and BxPC-3 were obtained from the American Type Culture Collection (Manassas, VA, USA); Calu-1, Calu-3, WI-38, MRC-5, T84, LS174T, SW1116 and AsPC-1 were obtained from the Shanghai Cell Bank of the Chinese Academy of Sciences (Shanghai, China). KRAS status identification by sequencing and limited genotyping in cancer cells was shown in Supplementary Table 2. Most of these cells were grown at 37 °C under a humidified 95:5 (%; v/v) mixture of air and CO2. SW480 and SW620 cells were grown without CO2. All of the cell lines were authenticated by short tandem repeat analysis.
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NSCLC and normal lung cell culture

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All cells were incubated at 37 °C in a humidified incubator with 5% CO2. NSCLC cell lines A549, H522, H596, H1975, HCC827, and PC9 were obtained from the American Type Culture Collection (Manassas, VA) and cultured in Roswell Park Memorial Institute (RPMI) 1640 Medium supplemented with 10% fetal bovine serum (FBS, Gibco), and penicillin-streptomycin. Lung normal cell lines MRC5 and WI38 were obtained from the Shanghai Cell Bank of the Chinese Academy of Sciences (Shanghai, China) and maintained in Eagle’s Minimum Essential Medium (EMEM) supplemented with 10% fetal bovine serum (FBS, Gibco) and penicillin-streptomycin (Supplementary Table 1). Cell lines were subjected to a mycoplasma detection test and authenticated by short tandem repeat (STR) profiling.
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