A micromethod for the activation of human blood lymphocytes in vitro

A micro culture system is described in which 2.5 X 10(4) human blood lymphocytes in aliquots of 100 mul 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 (10(4) responding cells in 40 mul 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.