The RKO, SW480, Lovo, HCT15, SW48, LS174T, SW620, LS180 and HCT116 colon cancer cell lines were used in this study and all were obtained from the American Type Culture Collection (Manassas, VA, USA). The cells were cultured in the recommended media (Dulbecco's modified Eagle's medium; Thermo Fisher Scientific, Inc., Waltham, MA, USA) for RKO, LS174T and LS180 and Roswell Park Memorial Institute-1640 medium (Sigma-Aldrich; Thermo Fisher Scientific, Inc.) for SW480, Lovo, HCT15, SW48, SW620 and HCT116 supplemented with 10% fetal bovine serum (Thermo Fisher Scientific, Inc.). Cells were maintained in a 37°C incubator in an atmosphere containing 5% CO2. Cells were regularly monitored using a light microscope and subcultured once they reached 80–90% confluency.
Ls180
Manufactured by Merck Group
Sourced in United States
The LS180 is a laboratory equipment product offered by the Merck Group. It serves as a general-purpose instrument for various laboratory applications. The LS180 is designed to perform specific functions without making claims about its intended use.
Automatically generated - may contain errors
Lab products found in correlation
Ls174t, by Merck Group (3 mentions)
Sw620, by American Type Culture Collection (3 mentions)
Ls180, by American Type Culture Collection (3 mentions)
Hct15, by Thermo Fisher Scientific (2 mentions)
Hct116, by American Type Culture Collection (2 mentions)
Hct116, by Thermo Fisher Scientific (2 mentions)
Hct15, by American Type Culture Collection (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (2 mentions)
Sw480, by American Type Culture Collection (2 mentions)
Sw480, by Thermo Fisher Scientific (2 mentions)
Ls174t, by American Type Culture Collection (2 mentions)
Sw620, by Thermo Fisher Scientific (2 mentions)
Ls180, by European Collection of Cell Cultures (2 mentions)
Streptomycin, by Merck Group (2 mentions)
Colo205, by American Type Culture Collection (1 mentions)
Sw837, by American Type Culture Collection (1 mentions)
Colo201, by American Type Culture Collection (1 mentions)
Rpmi 1640 medium, by Merck Group (1 mentions)
Colo205, by European Collection of Cell Cultures (1 mentions)
5 protocols using ls180
1
Culturing Colon Cancer Cell Lines
2
Colon Cancer Cell Line Culture
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The colon cancer cell lines (SW620, LoVo, SW480, HCT116, SW48, HCT15, RKO, SW837, COLO-201, COLO-205, LS174T and LS180) used in the present study were originally obtained from the American Type Culture Collection (Manassas, VA, USA). The cells were cultured in the recommended media (Dulbecco's modified Eagle's medium (Thermo Fisher Scientific, Inc., Waltham, MA, USA) for RKO, LS174T and LS180 and RPMI-1640 medium (Sigma-Aldrich; Merck KGaA, Darmstadt, Germany) for SW480, Lovo, HCT15, SW48, SW620 and HCT116 supplemented with 10% fetal bovine serum (Thermo Fisher Scientific, Inc.). The monolayer cells were maintained in a 37°C incubator with 5% CO2, observed regularly under a light microscope (magnification, ×40) and subcultured when they reached 80–90% confluency.
3
Overexpression of c-Src in Colon Cancer Cells
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COLO-205, SW48, SW480, LS180, LS174-T, HT-29, T84 and LoVo cells were purchased from the European Collection of Cell Cultures (ECACC). Cells in culture were maintained as a subconfluent monolayer in Dulbecco's Modified Eagle's medium supplied with non-essential amino acids (LS180 and LS174-T), Dulbecco's modified Eagle's medium-F12 nutrient mixture (T84), McCoy 5-A medium (HT-29), L15 Leivobitz medium (SW48 and SW480), Nutrient Mixture F12 HAM (LoVo) and RPMI 1640 (COLO-205) purchased from Sigma. Each cell line was grown under identical conditions, and cell culture medium supplements were provided according to the manufacturer's instructions. To ectopically overexpress c-Src kinase protein, subconfluent SW480 and T84 cells were transfected with 0.4 μg of pBABE-puro expression plasmid carrying cDNA from a c-Src gene. Stable clones of the T84 cell line were selected in F12/DMEM medium supplied with 0.5 μg of puromycin for 3 weeks. In the same way, stable clones of the SW480 cell line were selected in L15 Leivobitz medium supplied with 0.5 μg of puromycin. As a control, subconfluent SW480 and T84 cells were transfected with 0.4 μg of DNA containing an empty pBABE-puro expression plasmid. Positive clones were selected for protein expression measured by Western blotting. The entire set of transfected clones was used as a stable pool for transfection.
4
Proliferation Assays in LS180 Cells
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The human colon adenocarcinoma cell line LS180 was obtained from the European Collection of Cell Cultures (ECACC, Centre for Applied Microbiology and Research, Salisbury, UK). LS180 cells were grown in a 1:1 mixture of DMEM (Sigma-Aldrich, St. Louis, MO, USA) and Ham’s F-12 nutrient mixture (Sigma-Aldrich) supplemented with 10% foetal bovine serum (FBS) (Sigma-Aldrich), penicilin (100 U/mL) (Sigma-Aldrich) and streptomycin (100 μg/mL) (Sigma-Aldrich). Cells were maintained in a humidified atmosphere of 95% air and 5% CO2 at 37 °C.
The effects of extracts on cancer cells proliferation were determined by the MTT metabolism assay and BrdU incorporation immunoassay. The BrdU test was conducted using Cell Proliferation ELISA BrdU according to the manufacturer’s instructions (Roche Diagnostics GmbH, Penzberg, Germany). Both assays were performed in human adenocarcinoma cells (LS180). Details of the procedures are described elsewhere [52 (link)].
The effects of extracts on cancer cells proliferation were determined by the MTT metabolism assay and BrdU incorporation immunoassay. The BrdU test was conducted using Cell Proliferation ELISA BrdU according to the manufacturer’s instructions (Roche Diagnostics GmbH, Penzberg, Germany). Both assays were performed in human adenocarcinoma cells (LS180). Details of the procedures are described elsewhere [52 (link)].
5
Colorectal Cancer Cell Line Cultivation
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Four human colorectal cell lines representing different stages of tumor development were obtained from the American Type Culture Collection (ATCC, USA): HT29 (no. HTB-38), LS180 (no. CL-187), SW948 (no. CCL-237), and SW620 (no. CCL-227). HT29, LS180, SW948, and SW620 were grown in a 1:1 mixture of Dulbecco's Modified Eagle's Medium (DMEM)/F12 culture medium (Sigma, USA) supplemented with 10% fetal bovine serum (FBS; Gibco, UK), 100 U/mL penicillin, and 100 mg/mL streptomycin (Sigma). Human skin fibroblasts (HSFs) used as control cells were a primary cell line isolated from freshly excised fragments of human skin. 8 The HSF cell line was cultured in DMEM medium (Sigma) supplemented with 10% FBS and antibiotics. Human peripheral blood acute monocytic leukemia cells THP-1 (ATCC; no. TIB-202) were used as a model of human monocytes. The THP-1 cells were cultured in RPMI 1640 medium supplemented with 10% FBS and antibiotics.
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