Probing metabolic states of differentiating stem cells using two-photon FLIM

Scientific Reports
Aleksandra V MeleshinaElena V Zagaynova

Abstract

The ability of stem cells to differentiate into specialized cell types presents a number of opportunities for regenerative medicine, stem cell therapy and developmental biology. Because traditional assessments of stem cells are destructive, time consuming, and logistically intensive, the use of a non-invasive, label-free approach to study of cell differentiation provides a powerful tool for rapid, high-content characterization of cell and tissue cultures. Here, we elucidate the metabolic changes in MSCs during adipogenic differentiation, based on the fluorescence of the metabolic co-factors NADH, NADPH, and FAD using the methods of two-photon fluorescence microscopy combined with FLIM. To estimate the contribution of energy metabolism and lipogenesis in the observed changes of the metabolic profile, a separate analysis of NADH and NADPH is required. In our study we demonstrated, for the first time, an increased contribution of protein-bound NADPH in adipocytes that is associated with lipogenesis. The optical redox ratio FAD/NAD(P)H decreased during adipogenic differentiation, and that this was likely to be explained by the intensive biosynthesis of lipids and the enhanced NADPH production associated with this. Based on the data...Continue Reading

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Citations

Dec 23, 2017·Antioxidants & Redox Signaling·Olivia I Kolenc, Kyle P Quinn
Apr 5, 2018·Stem Cell Research & Therapy·Aleksandra V MeleshinaElena V Zagaynova
Oct 24, 2017·Journal of Biophotonics·Janna L MorrisonDouglas A Brooks
Jul 1, 2017·Journal of Biomedical Optics·Aisada Uchugonova
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Feb 23, 2020·PLoS Pathogens·Charles A CoomerSergi Padilla-Parra
May 19, 2020·Methods and Applications in Fluorescence·Deborah S BarkauskasMichael S Roberts
Jan 29, 2017·Stem Cell Research & Therapy·Aleksandra V MeleshinaElena V Zagaynova
Aug 15, 2019·Methods and Applications in Fluorescence·Svetlana A RodimovaElena V Zagaynova
Apr 17, 2020·Frontiers in Bioengineering and Biotechnology·Þóra SigmarsdóttirÓlafur E Sigurjónsson
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Jun 22, 2021·Frontiers in Cell and Developmental Biology·Thora Bjorg SigmarsdottirÓlafur Eysteinn Sigurjónsson
Jul 30, 2021·Nature Communications·Tongcheng QianMelissa C Skala

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Methods Mentioned

BETA
fluorescence microscopy
fluorescence

Software Mentioned

ImageJ
SPCImage
STATISTICA

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