A Micromethod for the Activation of Human Blood LymphocytesIn Vitro

Abstract

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.