title: Slow versus fast postthaw dilution for the recovery of lymphocytes in whole blood frozen in either dimethyl sulfoxide or glycerol.
abstract: It was previously shown by the authors that human whole blood was cryoprotected by dimethyl sulfoxide (DMSO) for use as inoculum in short-term lymphocyte cultures. That procedure utilized a fast postthaw dilution where the cryoprotective agent was diluted in two steps. In this paper it is shown that a five-step 'slow' dilution technique is much superior to the two-step 'fast' dilution procedure, which significantly revises our original report of 1972. This method has strikingly improved the degree of blastogenic response to phytohemagglutinin as measured by 3H-thymidine incorporation. The increase in amount of tritium incorporation was seen in cultures which were protected with either DMSO or glycerol. A similar effect was observed in unfrozen control cultures exposed to either cryoprotectant.
mesh_terms: D001793:Blood Preservation / Q000379:methods*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology*; D005615:Freezing; D005990:Glycerol / Q000494:pharmacology*; D006801:Humans; D008213:Lymphocyte Activation / Q000187:drug effects; D008214:Lymphocytes / Q000187:drug effects / Q000502:physiology*
chemical_list: D005990:Glycerol; D004121:Dimethyl Sulfoxide