NDLI: Nonlinear Regression Models for Determination of Nicotinamide Adenine Dinucleotide Content in Human Embryonic Stem Cells

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Nonlinear Regression Models for Determination of Nicotinamide Adenine Dinucleotide Content in Human Embryonic Stem Cells

Content Provider	Springer Nature Link
Author	Salykin, Anton Kuzmic, Petr Kyrylenko, Olga Musilova, Jindra Glatz, Zdenek Dvorak, Petr Kyrylenko, Sergiy
Copyright Year	2013
Abstract	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.
Starting Page	786
Ending Page	793
Page Count	8
File Format	PDF
ISSN	15508943
Journal	Stem Cell Reviews and Reports
Volume Number	9
Issue Number	6
e-ISSN	15586804
Language	English
Publisher	Springer US
Publisher Date	2013-07-06
Publisher Place	Boston
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	NAD Human embryonic stem cells hESC-derived fibroblasts Enzymatic cycling Nonlinear regression Biochemical assay Cell Biology
Content Type	Text
Resource Type	Article
Subject	Medicine

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Nonlinear regression models for determination of nicotinamide adenine dinucleotide content in human embryonic stem cells.

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