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@article{1126952,
author = {Salykin, Anton and Kuzmic, Petr and Kyrylenko, Olga and Musilová, Jindra and Glatz, Zdeněk and Dvořák, Petr and Kyrylenko, Sergiy},
article_location = {TOTOWA},
article_number = {6},
doi = {http://dx.doi.org/10.1007/s12015-013-9454-3},
keywords = {Biochemical assay; Enzymatic cycling; hESC-derived fibroblasts; Human embryonic stem cells; NAD; Nonlinear regression},
language = {eng},
issn = {1550-8943},
journal = {Stem Cell Reviews and Reports},
title = {Nonlinear Regression Models for Determination of Nicotinamide Adenine Dinucleotide Content in Human Embryonic Stem Cells},
volume = {9},
year = {2013}
}
TY - JOUR
ID - 1126952
AU - Salykin, Anton - Kuzmic, Petr - Kyrylenko, Olga - Musilová, Jindra - Glatz, Zdeněk - Dvořák, Petr - Kyrylenko, Sergiy
PY - 2013
TI - Nonlinear Regression Models for Determination of Nicotinamide Adenine Dinucleotide Content in Human Embryonic Stem Cells
JF - Stem Cell Reviews and Reports
VL - 9
IS - 6
SP - 786-793
EP - 786-793
PB - HUMANA PRESS INC
SN - 15508943
KW - Biochemical assay
KW - Enzymatic cycling
KW - hESC-derived fibroblasts
KW - Human embryonic stem cells
KW - NAD
KW - Nonlinear regression
N2 - Recent evidence suggests that energy metabolism contributes to molecular mechanisms controlling stem cell identity. For example, human embryonic stem cells (hESCs) receive their metabolic energy mostly via glycolysis rather than mitochondrial oxidative phosphorylation. This suggests a connection of metabolic homeostasis to stemness. Nicotinamide adenine dinucleotide (NAD) is an important cellular redox carrier and a cofactor for various metabolic pathways, including glycolysis. Therefore, accurate determination of NAD cellular levels and dynamics is of growing importance for understanding the physiology of stem cells. Conventional analytic methods for the determination of metabolite levels rely on linear calibration curves. However, in actual practice many two-enzyme cycling assays, such as the assay systems used in this work, display prominently nonlinear behavior. Here we present a diaphorase/lactate dehydrogenase NAD cycling assay optimized for hESCs, together with a mechanism-based, nonlinear regression models for the determination of NAD+, NADH, and total NAD. We also present experimental data on metabolic homeostasis of hESC under various physiological conditions. We show that NAD+/NADH ratio varies considerably with time in culture after routine change of medium, while the total NAD content undergoes relatively minor changes. In addition, we show that the NAD+/NADH ratio, as well as the total NAD levels, vary between stem cells and their differentiated counterparts. Importantly, the NAD+/NADH ratio was found to be substantially higher in hESC-derived fibroblasts versus hESCs. Overall, our nonlinear mathematical model is applicable to other enzymatic amplification systems.
ER -
SALYKIN, Anton, Petr KUZMIC, Olga KYRYLENKO, Jindra MUSILOVÁ, Zdeněk GLATZ, Petr DVOŘÁK a Sergiy KYRYLENKO. Nonlinear Regression Models for Determination of Nicotinamide Adenine Dinucleotide Content in Human Embryonic Stem Cells. \textit{Stem Cell Reviews and Reports}. TOTOWA: HUMANA PRESS INC, 2013, roč.~9, č.~6, s.~786-793. ISSN~1550-8943. Dostupné z: https://dx.doi.org/10.1007/s12015-013-9454-3.
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