As described previously (35 (link)), for ex vivo quantification of cytokine secretion, isolated lamina propria lymphocytes were washed with complete tissue culture media (RPMI, 10% FBS, 1% glutamine, and 1% 1:1 penicillin/streptomycin) and cultured in 1 mL of the same media in 24-well plates for 24 h at 37 °C, 5% carbon dioxide. The resulting supernatants were used to determine the amount of each cytokine using murine-specific ELISA kits (BD Biosciences) and a cytometrix bead array (CBA) flex set (BD Biosciences) relative to a standard curve according to the manufacturer’s recommendations.
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Ex Vivo Quantification of Cytokine Secretion
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Carbon dioxide
Cultured media
Cytokine
Elisa
Glutamine
Lamina propria
Lymphocytes
Murine
Penicillin
Secretion
Streptomycin
Tissue
Corresponding Organization :
Other organizations : Harvard University, Massachusetts General Hospital, Broad Institute
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Variable analysis
independent variables
- Isolated lamina propria lymphocytes were cultured
- Amount of each cytokine secreted
- Isolated lamina propria lymphocytes were washed with complete tissue culture media (RPMI, 10% FBS, 1% glutamine, and 1% 1:1 penicillin/streptomycin)
- Lymphocytes were cultured in 1 mL of the same media in 24-well plates for 24 h at 37 °C, 5% carbon dioxide
- Standard curve for ELISA and CBA flex set assays
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