After the introduction of mitochondria with a mixture of mutant and wild-type mitochondrial DNA (mtDNA) into a human rho degrees cell line (143B,206), Yoneda ct al, [Yoneda, M., Chomyn, A., Martinuzzi, A., Hurko, O, and Attardi, G, (1992) Proc. Natl. Acad. Sci. USA 89, 11164-11168] observed a shift in the proportion of the two mitochondrial genotypes in a number of cybrid clones, In every case where a shift was observed, there was an increase in the proportion of mutant mtDNA, By using the same cell line (143B,206 rho degrees), we also generated cybrids that were either stable in their mitochondrial genotype or showed an increase in the proportion of mutant mtDNA. However, temporal analysis of the same mutant mtDNA type in another rho degrees cell line revealed a quite distinct outcome. Those clones that showed a change shifted toward higher levels of wild-type rather than mutant mtDNA, These results indicate that the nuclear genetic background of the recipient (rho degrees) cell can influence the segregation of mutant and wild-type mitochondrial genomes in cell cybrids.