Herpesvirus saimiri subgroup-C strains transform non-neoplastic human T lymphocytes in vitro to antigen-independent continuous growth. The karyotypes of 8 of these T-cell lines, derived from peripheral blood or bone marrow, are analyzed here. In general, these lines showed a normal diploid karyotype, although one of them had acquired a clonal chromosomal rearrangement after 21 months in culture. Chromosomally aberrant cell clones were found more frequently in cell lines derived from blood lymphocytes of leukemia patients than in cultures from healthy donors. However, the observed aberrations, some of which were defined in detail using chromosome in situ suppression hybridization (CISS) with chromosome-specific recombinant DNA libraries, did not show any relation between the various cell lines nor to the respective types of leukemia.