To develop resources for large-scale production of human fibroblast interferon, we isolated, cryopreserved, and characterized 15 new strains of human diploid foreskin fibroblasts. Their life spans in vitro ranged from 52 to 72 population doublings. We based the selection of cell strains for mass interferon production on the number of population doublings during which consistently high yields of interferon were obtained after "superinduction" in roller bottles; our data show that aging in vitro leads to significant decline in amounts of interferon produced. In contrast, susceptibility to interferon remains largely unaffected by in vitro senescence. Karyotypic analysis indicated that the best interferon-producing strain, MLD (over 60,000 reference units/ml), has a translocation between chromosomes 5 and 15.