The processes in charge of what happens to human embryonic stem cells (hESCs) are thought to involve regulation by intrinsic and extracellular metabolic factors: amongst others, these include hormones and growth factors. A combination of technologies and approaches can help give more information about the mechanisms at play. High-resolution proton nuclear magnetic resonance (1H-NMR) spectroscopy, a powerful and non-invasive technique, is one of these. Using it together with statistical modelling methods can help characterise disturbances in metabolic pathways taking place during cellular events. This approach is called metabonomics. The 'Elucidation of Stem Cell Fate and Cell-type Characterization by 1H-NMR-based Metabonomics' (hESC metabonomics) project attempted to cover gaps in knowledge using this NMR-based metabonomics approach. The overall aim was to investigate metabolic changes related to hESC stem culture and directed cell differentiation. The use of two sets of serum samples in one-dimensional and two-dimensional NMR experiments proved very successful. The study revealed that NMR-acquired metabolic serum samples from chronic lymphocytic leukaemia patients can be used to quickly and accurately diagnose patients with an acute form of the disease in its early stages. Positive results were also obtained with blood metabolic profiling by NMR. This helped distinguish between patients suffering from cirrhosis and those with sub-clinical minimal hepatic encephalopathy. This means patients can now benefit from timely treatment, increasing their quality of life and likelihood of an improved outcome. Conducting hESC metabonomics' studies with culture media, which can provide a source of protein and metabolic factors, showed that NMR-based metabonomics offers a fast and accurate means of characterising hESC conditioning media. This should help researchers develop more effective culture conditions for increased production of hESCs in vitro to realise the potential of hESCs in a therapeutic context. The project managed to develop a robust model for studying metabolic changes during the differentiation of hESCs to motor neurons after a relevant protocol was established. Researchers were also able to identify certain metabolic pathways activated and deactivated during hESC differentiation to committed motor neurons.
Elucidation of Stem Cell Fate and Cell-type Characterization by 1H-NMR-based Metabonomics
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