I want to perform splenocyte restimulation culture for the estimation of Th1 and Th2 cytokines by ELISA. As each groups consists of 5 mice immunized with a test vaccine preparation; can splenocyte isolated from intra group mice be mixed together?
Yes you can; however, there should be enough splenocytes in each spleen to assess the response to your stimulus whether this is an antigen or other activating agent. The power of having 5 replicates each stimulated under 3 different conditions (i.e. stimulated with medium alone as a negative control; a mitogen such as concanavalin A as a positive control and your test agent) will allow you to get a clear result with solid statistical analysis from this single experiment. I would, therefore, strongly advise analyzing each spleen separately.
