HT29 cells (American Type Culture Collection, Manassas, VA, USA) were used to perform in vitro experiments. McCoy’s Medium Modified (Sigma-Aldrich, Madrid, Spain), supplemented with 100 U/mL penicillin, 100 µg/mL streptomycin, 2 mM L-glutamine and 10% inactivated FBS, was used to culture these cells. Depending on the experiment, HT29 cells were treated for 24 h with LPS 0.1 µg/mL, succinate 1 mM, the MEK inhibitor U0126 10 µM or an inflammatory cocktail containing TNF-α 25 ng/mL, IFN-γ 20 ng/mL, LPS 1 µg/mL.
Mccoy s medium modified
Manufactured by Merck Group
Sourced in United States
McCoy's Medium Modified is a cell culture medium that is used to support the growth and maintenance of various cell lines. It provides the necessary nutrients and components for cell proliferation and survival. The formulation is a modification of the original McCoy's 5A Medium, optimized for specific applications.
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Lab products found in correlation
Ht 29 cell, by American Type Culture Collection (2 mentions)
Xav939, by Merck Group (1 mentions)
Ht29 cells, by Merck Group (1 mentions)
Sodium pyruvate, by Merck Group (1 mentions)
Rpmi 1640, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Gentamicin sulfate, by Thermo Fisher Scientific (1 mentions)
Neutral red, by Merck Group (1 mentions)
Doxorubicin, by Merck Group (1 mentions)
Resazurin, by Merck Group (1 mentions)
Rhodamine 123 rho 123, by Merck Group (1 mentions)
Valencene, by Merck Group (1 mentions)
α humulene, by Merck Group (1 mentions)
β caryophyllene oxide, by Merck Group (1 mentions)
N 2 hydroxyethylpiperazine n 2 ethanesulfonic acid hepes buffer, by Merck Group (1 mentions)
Trans nerolidol, by Merck Group (1 mentions)
D glutamine, by Merck Group (1 mentions)
Penicillin streptomycin, by Merck Group (1 mentions)
Acridine orange, by Merck Group (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
4 protocols using mccoy s medium modified
1
Culturing and Treating HT29 Cells
2
Succinate and TGF-β Modulate HT29 Cells
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HT29 cells (American Type Culture Collection, VA, USA) were cultured in McCoy’s Medium Modified (Sigma-Aldrich, Madrid, Spain) supplemented with 10% inactivated FBS, 100 U/mL penicillin, 100 μg/mL streptomycin and 2 mM L-glutamine. HT29 cells were treated with different concentrations of succinate (0, 0.1, 0.5, 1 or 5 mM) or TGF-β (5 ng/mL) for 48 h or 7 days depending on the experiment. In the experiments of 7 days, the medium was changed every two days. In some cases, HT29 cells were treated with the inhibitor of the Wnt pathway, XAV939 (1 µM, Sigma-Aldrich, Madrid, Spain) for 48 h.
3
Cytotoxicity Evaluation of Sesquiterpenes
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McCoy’s modified medium, RPMI-1640, N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid (HEPES) buffer, sodium pyruvate, d -glutamine, resazurin, rhodamine 123 (RHO 123) and neutral red were supplied by Sigma-Aldrich (Prague, Czech Republic). Fetal bovine serum and gentamicin sulfate were purchased from Invitrogen (Carlsbad, CA, USA) and bovine serum albumin (BSA) from Fluka (Prague, Czech Republic). Doxorubicin and pure sesquiterpenes (α-humulene, β-caryophyllene oxide, trans-nerolidol, valencene) were supplied by Sigma-Aldrich. Stock solutions of sesquiterpenes were prepared in dimethyl sulfoxide (DMSO) and stored in the dark at 4 °C. All other chemicals used were of HPLC or analytical grade and provided by Sigma-Aldrich.
4
Biochemical Assays for Cell Metabolism
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Bovine serum albumin, thiobarbituric acid (TBA), phenazine methosulphate (PMS), nitroblue tetrazolium (NBT), 5,5-dithiobis(2-nitrobenzoic acid) (DTNB), 3-(4,5-dimethyl-2-thiaozolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), 2–7-diacetyl dichlorofluorescein (DCFH-DH), rhodamine 123 (Rh-123), acridine orange, ethidium bromide, heat inactivated fetal calf serum (FCS), minimum essential medium (MEM), McCoy’s modified medium, glutamine, penicillin-streptomycin, EDTA and trypsin were purchased from Sigma Chemicals Co. (USA).
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