HT-29 human epithelial cells were used for immunomodulation assays; either the parental lineage (HT-29, colon adenocarcinoma; ATCC HTB-38) or a lineage transfected with the secreted alkaline phosphatase (SEAP) reporter gene for NF-kB activation monitoring (HT-29/kb-seap-25) (Lakhdari et al., 2010 (link)). The reporter HT-29/kb-seap-25 cells were cultured in RPMI-Glutamine medium (Sigma-Aldrich), supplemented with 10% fetal bovine serum (Corning), 1% non-essential amino acids, 1% sodium pyruvate, 1% HEPES buffer (Thermofisher Scientific), and 1% penicillin-streptomycin (Lonza) according to Lakhdari et al. (2010) (link). The parental HT-29 cells were cultured in high-glucose DMEM medium (Dominique Dutscher) supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin (Do Carmo et al., 2017 (link)). For subcultures, cells were rinsed with DPBS (Thermofisher Scientific) and detached with a trypsin (0.05%) – EDTA (0.02%) solution (Sigma). Periodically, 100 μg/mL Zeocin (Invivogen) was applied to the HT-29/kb-seap-25 cell culture in order to maintain selective pressure on the cells containing the transfected plasmid.
Rpmi glutamine medium
Manufactured by Merck Group
RPMI-Glutamine medium is a cell culture medium developed by Merck Group. It is formulated to support the growth and maintenance of various mammalian cell lines in vitro. The medium contains L-glutamine, an essential amino acid required for cell metabolism and proliferation.
Automatically generated - may contain errors
Lab products found in correlation
Dulbecco phosphate buffered saline (dpbs), by Thermo Fisher Scientific (4 mentions)
Fetal bovine serum (fbs), by Dutscher (2 mentions)
Penicillin streptomycin, by Dutscher (2 mentions)
Hepes buffer, by Thermo Fisher Scientific (2 mentions)
High glucose dmem medium, by Dutscher (2 mentions)
Zeocin, by InvivoGen (2 mentions)
Penicillin streptomycin, by Lonza (2 mentions)
Fetal bovine serum (fbs), by Corning (2 mentions)
Trypsin, by Merck Group (1 mentions)
Ethylene diamine tetraacetic acid (edta), by Merck Group (1 mentions)
Trypsin 0.05 edta 0 02 solution, by Merck Group (1 mentions)
2 protocols using rpmi glutamine medium
1
Immunomodulation Assays using HT-29 Cells
2
HT-29 Epithelial Cell Immunomodulation
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HT-29 human epithelial cells were used for immunomodulation assays; either the parental lineage (HT-29, colon adenocarcinoma; ATCC HTB-38) or a lineage transfected with the secreted alkaline phosphatase (SEAP) reporter gene for NF-kB activation monitoring (HT-29/kb-seap-25) (Lakhdari et al., 2010) . The reporter HT-29/kb-seap-25 cells were cultured in RPMI-Glutamine medium (Sigma-Aldrich), supplemented with 10% fetal bovine serum (Corning), 1% non-essential amino acids, 1% sodium pyruvate, 1% HEPES buffer (Thermofisher Scientific), and 1% penicillin-streptomycin (Lonza) according to Lakhdari et al. (2010) . The parental HT-29 cells were cultured in high-glucose DMEM medium (Dominique Dutscher) supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin (Do Carmo et al., 2017) . For subcultures, cells were rinsed with DPBS (Thermofisher Scientific) and detached with a trypsin (0.05%) -EDTA (0.02%) solution (Sigma). Periodically, 100 µg/mL Zeocin (Invivogen) was applied to the HT-29/kb-seap-25 cell culture in order to maintain selective pressure on the cells containing the transfected plasmid.
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