Demem high glucose 4.5 g l medium

Mouse embryonic fibroblasts from mtDNA mutator mice were generated as previously described [64] . Wildtype (WT) and mutator MEFs were cultured in DEMEM High Glucose (4.5 g/l) medium (without Sodium pyruvate) (ThermoFisher Scientific, Rockford, USA) supplemented with 10 % FBS, 1 % penicilin/streptomycin at 37 °C and 5 % CO 2 . MEFs isolated from four individual mtDNA mutator and three individual WT mice were used as different biological replicates [25] (link). Primary skin fibroblasts were cultured in DEMEM High Glucose (4.5 g/l) medium (without sodium pyruvate) (ThermoFisher Scientific, Rockford, USA) supplemented with 10 % FBS, 1 % penicilin/streptomycin and 200 µM uridine at 37 °C and 5 % CO 2 . Cells were grown to approx. 70 % confluency before passaging them into new flasks or using them for experiments. At least three different passages of the individual patients' lines were used as biological replicates. Primary human lung fibroblasts were cultured in DEMEM High Glucose (4.5 g/l) medium (without sodium pyruvate) (ThermoFisher Scientific, Rockford, USA) supplemented with 10 % FBS, 1 % penicilin/streptomycin, 2 mM glutamine (ThermoFisher), 2 ng/ml β-FGF (ThermoFisher), 0.5 ng/ml EGF (Sigma-Aldrich) and 5 µg/ml insulin (ThermoFisher).