Transcriptome changes accompany mitochondrial membrane potential heterogeneity in mouse embryonic and induced pluripotent stem cells

Stem cells are uniquely dependent on both oxidative phosphorylation and glycolysis to maintain pluripotency and drive differentiation. Here, we identified that stem cells sorted by their inherent differences in mitochondrial activity exhibited significantly altered germline differentiation potential. Stem cells with lower average mitochondrial membrane potential were less proliferative and generated less ROS and ATP despite having a similar mitochondrial DNA copy number as cells with higher membrane potential. We demonstrate that pluripotent stem cells exhibit a large range of mitochondrial activity from an otherwise homogenous population, and that they are dependent on this activity to drive differentiation. Mouse embryonic and induced pluripotent stem cells from a C57BL/6 background were stained with mitochondrial membrane potential dye tetramethylrhodamine methyl ester (TMRM) and sorted by fluorescence activated cell sorting into populations of low or high mitochondrial membrane potential. RNA was extracted from each population and analyzed by Affymetrix arrays.