Identification of key regulatory metabolites in hESCs by MALDI-TOF-MS

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Authors

SALYKIN Anton AMATO Filippo DVOŘÁK Petr ROTREKL Vladimír HAVEL Josef

Year of publication 2014
Type Appeared in Conference without Proceedings
MU Faculty or unit

Faculty of Science

Citation
Description The ability of human embryonic stem cells (hESC) to differentiate into any cell type of human body presents a tool for regenerative medicine raising the need for advanced cultivation and differentiation protocols. Metabolic homeostasis in stem cells is shown to be different from that in somatic cells. To meet their energetic demands stem cells rely mostly on glycolysis rather than on oxidative phosphorylation. Energy metabolism contributes to molecular mechanisms controlling stem cell identity, selfrenewal and differentiation. In this context, clear understanding of how cell fate decisions and metabolic processes are intertwined is essential for identification of key regulatory nodes capable of differential stem cell fate choice regulation. Matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI TOF MS) has been proved to be a powerful tool to study proteomic and metabolic fingerprints of both bacterial and mammalian cell lines. The technique with optimized stem cell extraction protocol allowed for identification of more than 20 individual metabolites contributing to the dynamic changes in human pluripotent stem cell metabolism. By employing advanced analysis we were able to identify the key metabolites that contribute to the maximum changes in the stem cell metabolism upon varying physiological conditions. Our data shed light on the plasticity of metabolic homeostasis in human pluripotent stem cells during cultivation in vitro opening the space for development of more specific cultivation protocols for translational medicine.
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