I plan on evaluating PBMC supernatants with the Biolgend Legendplex bead array kit (13 inflammation panel - IL-2, IL-4, IL-5, IL-6, IL-9, IL-10, IL-13, IL-17A, IL-17F, IL-21, IL-22, IFN-γ and TNF-α).
Do I need to have separate wells/plates for each time point collection?
Would the longer incubation periods (48 and 72hrs) require fresh medium added to the PBMCs to keep them going?
I rather recommend you to perform preliminary experiments.
1) Do you know the best condition of PBMC and bacterial cell ratio in your culture system? For example 1x10
shorter period is better (6-8 hrs) especially if you work with pathogenic bacteria
Thank you Toshimitsu, I have performed some preliminary experiments measuring TNF-a only using 2x10^6 PBMC to 0.5mg/ml of bacterial cells (this cells are killed and lyophilized then resuspended).
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