Cryopreservation of Human Lymphocytes: A Brief Review and Evaluation of an Automated Liquid Nitrogen Freezer

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Abstract
Successful cryopreservation of human lymphocytes was previously described. Cryopreserved lymphocytes are useful for a variety of in vitro immunologic studies. In this study, the applicability and/or advantages of using a programmable freezing system was determined and glycerol vs. dimethyl sulfoxide (DMSO) was compared at varying concentrations on post-thaw viability, E[erythrocyte]-rosetting and immunoglobulin fluorescence. Prefreeze T [thymus-derived] and B [bone marrow-derived] lymphocyte percentages were determined. Cells were then frozen in varying concentrations of glycerol and DMSO. Optimum cryoprotectant type and concentration was determined. Lymphocytes from 7 individuals were frozen by the batch method in a mechanical freezer and with the automated liquid nitrogen injection system. Data on post-thaw T and B percentages and viability revealed 10% DMSO and liquid nitrogen control freezing method at 1.degree. C/min as the best conditions for lymphocyte preservation, as reflected by post-thaw in vitro testing.