The current study provides a multimodal molecular map across the entire replicative lifespan of primary human fibroblast cell lines from n=6 healthy male and female donors and n=3 donors with lifespan-altering mitochondrial disease caused by mutations in the SURF1 gene. These data include cytological (cell size, morphology), bioenergetic (energy expenditure, derived ATP synthesis rates from Seahorse), secreted proteins, telomere length, and whole-genome sequencing (WGS) data. These datasets can be integrated with other omic measures collected in parallel along the cellular lifespan, including gene expression (RNA sequencing) and DNA methylation (GSE179849).This dataset also includes experimental manipulations with treatments targeting oxidative phosphorylation (OXPHOS) and glycolysis, and glucocorticoid signaling, providing an opportunity to examine the influence of stress and bioenergetics on human aging biology. All processed data can be accessed and browsed at our webtool: https://columbia-picard.shinyapps.io/shinyapp-Lifespan_Study/. A detailed description of the study and associated methods can be found at: https://www.biorxiv.org/content/10.1101/2021.11.12.468448