Stem cells hold outstanding potential to model and treat disease and are valuable tools in pharmacology and toxicology. Characterization of stem cells and derivatives still poses many challenges to ensure safe, efficacious, and reliable therapies. Regulatory agencies have defined key mandatory attributes related to identity, purity, sterility, and genomic integrity, however robust analytics to determine cell's potency are still a major challenge, in most cases assessed case-by-case. Importantly, the application of high-throughput ‘omic tools is opening new perspectives on stem cell's research and development. Here, analytical methodologies currently employed to characterize stem cells’ quality attributes are discussed, with special focus on ‘omics as relevant tools for definition of cell's mechanism of action, and for potency assay development and assessment.