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3 protocols using ccd841 con

1

Culturing Human Colon Cell Lines

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Two human CRC cell lines, DLD-1 and HCT116, as well as a normal colon epithelial cell line, CCD841 CoN, were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). According to company’s instructions, DLD-1, HCT116 and CCD841 CoN cells were cultured in the RPMI-1640 medium (Sigma-Aldrich, St. Louis, MO, USA, catalog #R8758), McCoy’s 5a Modified Medium (Life Technologies, Carlsbad, CA, USA, catalog #16600-082) and Eagle’s minimum essential medium (ATCC, catalog #30-2003), respectively. Cells were maintained at 37 °C in a humidified incubator with 5% CO2/95% air.
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2

Colorectal Cancer Cell Lines Cultivation

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Three colorectal cancer cell lines were used in the experiments: HCA-2 (Duke C), LS 174T (Duke B), and SW1116 (Duke A), together with a normal cell line, CCD 841 CoN. All cell lines were supplied by ATCC (American Type Culture Collection ATCC®, Old Town Manassas, VA, USA). To ensure optimal conditions for cell growth, appropriate culture media were used: Eagle’s minimum essential medium (EMEM) (ATCC 30-2003) for CCD 841CoN and LS 174T cell lines, and Dulbecco’s modified Eagle’s medium/Nutrient Mixture F-12 Ham (DMEM), Sigma-Aldrich D8437). For SW1116 and HCA-2 cell lines, both media were supplemented with 10% foetal bovine serum ((FBS), ATCC 30-2020) and 1% penicillin–streptomycin–neomycin stabilised solution (Sigma-Aldrich P4083).
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3

Cell Line Characterization for Research

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In the presented project, four cell lines were used: two carcinoma, HCT116 and H460, and two normal, CCD 841 CoN and MRC-5. All cell lines were purchased from the American Type Culture Collection (Manassas, VA, ATCC) and were tested negatively for mycoplasma using the Universal Mycoplasma Detection Kit (ATCC). Colorectal carcinoma HCT116 cells were maintained in McCoy’s 5A medium (Merck/Sigma-Aldrich, Darmstadt, Germany), and non-small-cell lung carcinoma H460 cells were maintained in RPMI 1640 medium (Merck/Sigma-Aldrich, Germany). Both media were supplemented with 10% fetal bovine serum (FBS; Biowest, Riverside, MO, USA), 100 μg/mL streptomycin and 100 unit/mL of penicillin. Human colon epithelial CCD 841 CoN cells and human lung fibroblast MRC-5 cells were maintained in Eagle’s Minimal Essential Medium (MEM; Merck/Sigma-Aldrich) supplemented with 10% FBS without antibiotics. All cells were incubated in 5% CO2 atmosphere at 37 °C. The experiments were performed with cells in the exponential phase of growth.
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