Human influenza viruses used for vaccine production have previously been adapted to grow in eggs. During egg adaptation, variants are selected and we have observed that more than one variant may derive in a single egg resulting in a mixed population. We have now investigated the extent of heterogeneity, due to host cell selection, of virus strains used for the manufacture of influenza vaccine for the 1991/1992 and 1992/1993 seasons. The A(H1N1) vaccine virus was homogeneous with respect to substitutions in the haemagglutinin deriving from egg adaptation. However, two A(H3N2) vaccine strains and the influenza B component, B/Yamagata/16/88, consisted of mixed populations, apparently due to their cultivation in eggs. The individual variants within B/Yamagata were isolated and found to be antigenically distinct. The ratios of these variants within different manufacturers' seed stocks varied to the extent that vaccine derived from them could be distinguished antigenically. Furthermore, derivation of high-growth reassortants from the A(H3N2) strains which involves passaging at limit dilution did not necessarily lead to a homogeneous virus population. The significance of these findings for the efficacy of vaccine is not known at present.