A strategy has been developed to detect and characterize certain heritable genomic alterations that occur as cells proliferate in vitro. Multiple subclones of cells were isolated from two clonal lymphoblastoid cell lines--one from a boy with Bloom's syndrome (BS), a cancer-predisposing condition known to feature excessive somatic mutation, the other from a normal man. The DNAs from the cell lines were hybridized to a panel of probes that can detect restriction fragment length polymorphisms, and the patterns of polymorphism in the primary clones were compared with that in each of the secondary clones. In one of the BS secondary clones three loci, positioned distally on the long arm of chromosome 3 and that are heterozygous in the donor and all other cell lines derived from the primary clone, had lost heterozygosity and apparently had become homozygous; in contrast, heterozygous loci more proximal on 3q had retained their heterozygosity, as had those on 3p. Taking into account the pattern of chromosome instability uniquely characteristic of BS, the most plausible explanation for the alterations in the altered clone is that somatic recombination had occurred in vitro, via homologous chromatid interchange. Such spontaneous recombinational events in nonneoplastic, nonmutagenized cells may contribute to the high cancer incidence in BS and, by analogy, to cancer that arises in the general population.