mtDNA mutants cybrid cell line construction Rhoo cell culture
MtDNA mutant cell line cybrids construction:
mtDNA sequencing generates a panel of new variants unknown in databases. To test their involvement in a bioenergetic defect, we construct transmitochondrial cybrid cell lines. This is achieved by fusing Rho0 143B TK- cells (devoid of mtDNA) with enucleated fibroblasts or other tissue patient cells. Transmitochondrial cybrids are then selected by bromouridine addition. Bioenergetic analysis on cybrid cells are conducted and if the primary defect is also observed in those cells, we can conclude on the involvement of the identified mtDNA variant in the clinical phenotype.