A Micromethod for the Activation of Human Blood Lymphocytes In Vitro

A micro culture system is described in which 2.5 × 10⁴ human blood lymphocytes in aliquots of 100 μL are stimulated by PHA, Pokeweed, “Varidase” antigen, allogeneic small lymphocytes or mitomycin-C-treated allogeneic LCL cells. Careful regulation of the pH by a combination of bicarbonate and MOPS buffers seems to be important for detecting a response to weak stimuli. High and reproducible levels of activation by powerful stimuli (PHA and LCL cells) can be recorded from even smaller cultures (10⁴ responding cells in 40 μL aliquots). The technique allows large numbers of replicate cultures to be set up from a single blood sample so that the time course and/or dose-response relationships can be examined for a range of differen mitogens.