supernatant harvesting - (Nov/23/2011 )

When doing timepoint analysis is it necessary to use a different plate of cells for each timepoint? Would not doing so (taking media from a single well at each timepoint) result in an effect where the release of cytokine is more concentrated at later timepoints? It's an adherent cell line.

It really depends on how close your time points are. If it is a matter of minutes (5 - 30 min), I'd say go with separate cultures since you don't want to factor in discrepancies due to removing the plate from the incubator too often to affect cytokine release (variable CO2 and temperatures). Even if it's 1 hour intervals, I personally wouldn't repeatedly remove the treatment plate from the incubator.