Mccoy s 5a medium

Human tissue samples of CRC and matched non-tumor tissues (ie, at least 5 cm away from the tumor edge) were obtained from patients who had undergone surgical treatment at the Third Xiangya Hospital of Central South University. All examined tumor tissues were sporadic adenocarcinomas. The use of tissues for this study was approved by the ethics committee of the Third Xiangya Hospital of Central South University, and the study was conducted in accordance with the Declaration of Helsinki. Before using these clinical materials for research purposes, all patients signed an informed consent. None of these patients received any preoperative chemotherapy or radiotherapy. The human CRC cell lines (HCT116 and HT29) were purchased from commercial sources and cultured in the Central Laboratory of the Third Xiangya Hospital of Central South University. HCT116 and HT29 cells were cultured in McCoy’s 5A medium (KeyGEN BioTECH, Nanjing, China) and Supplemented with 10% FBS (BioInd, Kibbutz Beit Haemek, Israel) in a humidified atmosphere with 5% CO₂ at 37°C.

We obtained human colorectal cancer cell lines either from our lab, the Chinese Academy of Sciences, or the Shanghai Institute of Biological Sciences. Dulbecco’s Modified Eagle’s Media (Gibco, Thermo Fisher Scientific, Inc.) was used to treat RKO, SW480, and SW620 cells, whereas McCoy’s 5A medium was used to treat HCT116 cells (Nanjing KeyGen Biotech Co., Ltd.). We added 10% foetal bovine serum (Gibco, Thermo Fisher Scientific, Inc.) and 1% penicillin-streptomycin to the media as supplements. To continue exponential development, subculture was performed every two to three days at 37°C in a humidified environment with 5% CO₂ and 95% air.

Human SW480 and HCT116 CRC cell lines were obtained from the Cell Center of Xiangya School of Medicine, Central South University (Hunan, China). SW480 and HCT116 were maintained in L-15 and McCoy’s 5A Medium (KeyGen Biotech, Nanjing, China) with 10% FBS (Biological Industries, Beit Haemek, Israel). All cells were maintained in 1% penicillin/streptomycin and cultured at 37°C in a humidified incubator with 5% CO₂.

Human CRC cell lines (SW620 and SW480) were cultured in L15 medium (KeyGEN BioTECH, Nanjing, China) supplemented with 10% fetal bovine serum (FBS) (Biological Industries, Israel), and HCT116 and HT-29 cells were cultured in McCoy‘s 5A medium (KeyGEN BioTECH, Nanjing, China) supplemented with 10% FBS. Cells were grown in a 5% CO₂ cell culture incubator at 37 °C. Clinical samples were obtained from patients treated at the Third Xiangya Hospital of Central South University (Hunan, China) under informed consent and approval by the Ethics Committee of Central South University (more details about the clinical samples can be found in Table 1).