Human Embryonic Stem Cells (hESCs) harbour the extraordinary ability to self-renew and differentiate into cells of all tissue types and thus, hold great promise for the treatment of a range of diseases. Mechanisms controlling hESC fate are yet to be elucidated but are believed to involve regulation by intrinsic and extracellular metabolic factors including hormones, growth factors and small cell-permeable molecules. High resolution proton Nuclear Magnetic Resonance (1H-NMR) spectroscopy is a powerful, non-invasive technique that can be used in conjunction with statistical modeling methods to characterize perturbations in metabolic pathways that occur during cellular events. This approach, termed ‘metabonomics’, can also be used to acquire a metabolic fingerprint of a cell at a particular time point or state. Such information can be used for better comprehending underlying cellular events as well as for cell-type classification. In this project, NMR-based metabonomics will be employed to realise two main objectives; 1) identification and characterisation of metabolic factors involved in hESC differentiation, and 2) cell-type classification of differentiated hESCs. These objectives are closely interrelated as the elucidation of factors controlling stem cell fate would provide the means for “directed cellular differentiation” and prove invaluable for regenerative medicine and drug discovery applications.