Molecular characterization of mtDNA depleted and repleted NT2 cell lines

Transmitochondrial cytoplasmic hybrids (cybrids) enable functional assessment of mitochondrial DNA (mtDNA)-encoded proteins. Cybrid production often utilizes cell lines depleted of endogenous mtDNA (ρ0 cells), and a number of suitable ρ0 cell lines exist for this purpose. We now provide molecular data characterizing an NT2 human teratocarcinoma ρ0 cell line, as well as NT2 cybrid derivatives. NT2 ρ0 cells contained no detectable mtDNA on a sensitive PCR assay. Eight weeks after exogenous mtDNA transfer cybrids showed no evidence of endogenous mtDNA reversion, and heteroplasmic ratios of a single nucleotide substitution roughly reflected that of the blood samples used to repopulate their mtDNA.