Two human lymphoblastoid cell lineages derived from the same parental line exhibit markedly different survival and mutational responses to X-irradiation, but not to chemical point mutagens. WI-L2-NS (ATCC CRL 8155) and TK6 (ATCC CRL 8015) both are derived from the original WI-L2 isolate described by Levy et al. (1968). Both lines are near diploid with stable and indistinguishable karyotypes (47, X, Y 13 +). However, differences in the extent of heterozygosity of chromosome 17 RFLP markers have been detected in these lines. Relative to TK6, WI-L2-NS and several cell lines subsequently derived from it exhibit enhanced survival after X-ray treatment. This is due partly to a more pronounced shoulder in the dose response curve for WI-L2-NS and partly to a higher D0 than is observed in TK6. X-ray-induced mutant frequencies also are markedly different. At the hprt locus, the overall magnitude of the response is similar in the two cell lines. However, in TK6, a linear equation appears to be the best fit to the data, as compared to a linear quadratic curve for WI-L2-NS. Induced mutant frequencies at the tk locus in heterozygotes derived from WI-L2-NS are 20-50-fold higher than those seen in TK6 and tk heterozygous derivatives of TK6. Analysis of the mutability of the two tk alleles in various tk heterozygotes of WI-L2-NS reveals a similar pattern to that described previously in heterozygotes derived from TK6; 3 times as many mutants were recovered from one tk allele than the other. A possible explanation for the higher survival and induced mutant frequencies seen in WI-L2-NS and its derivatives is the presence in these lines of an error prone repair system not functioning in TK6.