Hi, did you try to cut the lympnode into little peaces before pressing throug a maize into RPMI? If not, the cells will be squeezed and die. If you store the T-cells more than 24 hours you shoud use cRPMI (10%fetal cow serum and 1% Penstrep and 1% glucose). Do not use NaOH (cells may burst) use RPMI or cRPMI for restoring. regards Susanne