Factors Influencing the Stimulation of Human Lymphocytes by Antigen during Culture in vitro

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Abstract
Studies were undertaken to determine culture conditions which supported optimal survival of lymphocytes in vitro and maximal response to PPD. Optimal conditions included: (a) lymphocyte preparations not completely devoid of other leukocytes; (b) a final concentration of 1 million lymphocytes/ml; (c) a reaction mixture containing 1 μg PPD/ml and 20% pooled normal human serum; (d) incubation for 5 days at 37°C; (e) an environment of 5% CO2 in air, and (f) incubation at 5°C from the horizontal in sealed round-bottom culture tubes. A satisfactory substitute for whole human serum was not found. Despite control of these conditions considerable variation in response was encountered.

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