Accessory cell requirements in mitogen-induced rabbitt lymphocyte proliferation in an improved tissue culture system

Summary The impact of an improved culture medium (IMDM⁺), consisting of Iscove’s modified Dulbecco’s medium supplemented with albumin, transferrin, insulin, zinc, 2-mercapthoethanel, and 0.1% fetal bovine serum was investigated in phytohemagglutinin (PHA)-induced rabbit T cell proliferation. At the number of 2×10⁵ cells/well purified T lymphocytes cultured in IMDM⁺ responded 3 to 10 times better than lymphocytes cultured in serum-supplemented RPMI 1640 medium. In these conditions, PHA-induced proliferation seemed not to require the presence of accessory cells. However, at lower cell numbers, T cell proliferation was more efficient when calculated on a per cell basis. At these low cell numbers, optimal proliferation required accessory cells like macrophages or dendritic cells. The appraent absence of this requirement for accessory cells at high T cell concentrations may be explained by the contribution of contaminating macrophages and dendritic cells in the purified T cell fractions.