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A simple and inexpensive method for the freezing of human blood lymphocytes at -80 C is described. Lymphocytes were frozen in either polystyrene tubes or polyolefin bags. Optimal recovery of cells was obtained when 5% DMSO and 62% plasma were used as cryoprotectants. Viable cell recovery, as assessed by response to phyto hemagglutinin (PHA) and pokeweed mitogen (PWM), was greater than 80% after 60 days of storage. In addition, percentages of T and B lymphocytes were essentially unchanged. The excellent viable cell recovery in polyolefin bags suggests that it is possible to cryopreserve large numbers of lymphocytes in a single container.
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