Established human CRC cell lines (HCT116, LS180, COLO205, HT29 and SW480) were purchased from European Collection of Cell Cultures (ECACC, Salisbury, UK). HCT116, LS180 and COLO205 were maintained in RPMI-1640 supplemented with 10% FBS, GlutaMAX-I, non-essential amino acids (NEAA), 100 mM sodium pyruvate, 10 mM HEPES (all from GIBCO) and 50 mM 2-mercaptoethanol (Sigma–Aldrich, St. Louis, MO). HT29 was maintained in McCoy’s 5A medium (Sigma) supplemented with 10% FBS and GlutaMAX-I. SW480 were cultured in L-15 Medium (Leibovitz) (Sigma–Aldrich) supplemented with 10% FBS and GlutaMAX-I. Kanamycin sulfate (GIBCO) was included with all media. Absence of mycoplasma contamination in cultured cells was verified by PCR testing prior to investigation.
Colo205
Manufactured by European Collection of Cell Cultures
COLO205 is a continuous cell line derived from a human colon adenocarcinoma. It is a commonly used in vitro model for colorectal cancer research.
Automatically generated - may contain errors
Lab products found in correlation
Ls180, by European Collection of Cell Cultures (3 mentions)
Sw480, by European Collection of Cell Cultures (2 mentions)
Mccoy s 5a medium, by Merck Group (2 mentions)
Ht 29, by Merck Group (2 mentions)
Rpmi 1640, by Thermo Fisher Scientific (2 mentions)
Kanamycin sulfate, by Thermo Fisher Scientific (1 mentions)
Colo205, by Thermo Fisher Scientific (1 mentions)
Ls180, by Thermo Fisher Scientific (1 mentions)
Hepes, by Thermo Fisher Scientific (1 mentions)
Hct116, by Thermo Fisher Scientific (1 mentions)
2 mercaptoethanol, by Merck Group (1 mentions)
Hct116, by European Collection of Cell Cultures (1 mentions)
Nutrient mixture f 12 ham, by Merck Group (1 mentions)
Ls174t, by Merck Group (1 mentions)
Ls180, by Merck Group (1 mentions)
Ls174 t, by European Collection of Cell Cultures (1 mentions)
Kanamycin, by Thermo Fisher Scientific (1 mentions)
α mem, by Thermo Fisher Scientific (1 mentions)
Ebm 2 medium, by Lonza (1 mentions)
Glutamax 1, by Thermo Fisher Scientific (1 mentions)
3 protocols using colo205
1
Culturing Human Colorectal Cancer Cell Lines
2
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.
3
Culturing Human CRC Cell Lines
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Human CRC cell lines LS180, COLO205 and HT29, purchased from the European Collection of Cell Cultures, were maintained in RPMI 1640 (GIBCO) or, for HT29, McCoy's 5A medium (Sigma-Aldrich), supplemented with 10% fetal bovine serum, GlutaMAX-I and kanamycin (GIBCO). HMEC cells (provided by Professor T Resink, University of Basel) were cultured in EBM-2 medium (Lonza). Tumour-associated stromal cells (TASC) were expanded from CRC samples in α-MEM (GIBCO) supplemented with 10% fetal bovine serum and 5 ng/mL FGF-2 (R&D Systems).
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