I want to harvest RNA from CD8 T cells from mice spleens. In order to minimize RNA degradation I intend to freeze the spleens in liquid nitrogen before homogenization. Will this affect CD8 T cell isolation?
Hi! I haven't practically tried this myself, but I would suppose the freeze-thaw will effect immune cell isolation from spleens. The effect will depend upon exact conditions and time. In our lab we use a proteomic stabilizer buffer to minimise the effect of -80C freezing on PBMCs and give us optimal immune cell count as well as expression at a later time.
Thus, it might be better to extract T cells from fresh spleens and may be freeze isolated cells at -80C with a suitable buffer to allow for RNA processing at a later timepoint? Would be recommended to confirm with a small pilot first.