Protein supplements, mitogens and antibiotics in lymphoblast transformation and cytogenetic analysis

The in vitro effects of four protein supplements, foetal calf serum (FCS), gammaglobulin-depleted FCS (FCS-A), human serum albumin (HSA) and autologous serum (AS), and of four antibiotics on lymphocyte blast transformation and on lymphocyte proliferation for cytogenetic studies, were evaluated. 3H-thymidine incorporation was measured in the lymphocytes obtained from sixteen healthy adults and stimulated with phytohaemagglutinin (PHA), concanavalin A (Con A), pokeweed mitogen (PWM), PWM + PHA and PWM + Con A. The most efficient protein supplement was AS, although great variability was observed between subjects. HSA came second, while FCS and FCS-A were the least effective. The presence of immunoglobulins did not modify mitogen-induced lymphocyte responses. In unsupplemented media, peak responses to PHA and Con A occurred at lower mitogen concentrations. Furthermore, the addition of the antibiotics tested did not impede blast transformation whether or not a protein supplement was present. This work also showed that for chromosome analysis, protein supplements are necessary for optimum whole-blood lymphocyte cultures and that HSA can replace FCS.