Abstract
Cellular metabolism contributes to cell fate decisions. Bioenergetic profiling can therefore provide considerable insights into cellular identity and specification. Given the current importance of human pluripotent stem cells (hPSCs) for biomedical applications, assessing the bioenergetic properties of hPSCs and derivatives can unveil relevant mechanisms in the context of development biology and molecular disease modeling. Here, we describe a method to facilitate bioenergetic profiling of hPSCs in a reproducible and scalable manner. After simultaneous assessment of mitochondrial respiration and glycolytic capacity using Seahorse XFe96 Analyzer, we measure lactate concentration in the cellular media. Finally, we normalize the values based on DNA amount. We describe the procedures with specific requirements related to hPSCs . However, the same protocol can be easily adapted to other cell types, including differentiated progenies from hPSCs .
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References
Evans MJ, Kaufman MH (1981) Establishment in culture of pluripotential cells from mouse embryos. Nature 292(5819):154–156
Thomson JA, Itskovitz-Eldor J, Shapiro SS, Waknitz MA, Swiergiel JJ, Marshall VS et al (1998) Embryonic stem cell lines derived from human blastocysts. Science 282(5391):1145–1147
Takahashi K, Yamanaka S (2006) Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 26(4):663–676
Takahashi K, Tanabe K, Ohnuki M, Narita M, Ichisaka T, Tomoda K et al (2007) Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell 131(5):861–872
Lisowski P, Kannan P, Mlody B, Prigione A (2018) Mitochondria and the dynamic control of stem cell homeostasis. EMBO Rep 19(5):4067162
Xu X, Duan S, Yi F, Ocampo A, Liu GH, Izpisua Belmonte JC (2013) Mitochondrial regulation in pluripotent stem cells. Cell Metab 18(3):325–332
Lorenz C, Lesimple P, Bukowiecki R, Zink A, Inak G, Mlody B et al (2017) Human iPSC-derived neural progenitors are an effective drug discovery model for neurological mtDNA disorders. Cell Stem Cell 20(5):659–74 e9
Zheng X, Boyer L, Jin M, Mertens J, Kim Y, Ma L et al (2016) Metabolic reprogramming during neuronal differentiation from aerobic glycolysis to neuronal oxidative phosphorylation. Elife 5
Avior Y, Sagi I, Benvenisty N (2016) Pluripotent stem cells in disease modelling and drug discovery. Nat Rev Mol Cell Biol 17(3):170–182
Acknowledgments
The authors declare no competing financial or commercial interests and acknowledge support from the German Federal Ministry of Education and Research (BMBF) (#AZ.031A318 and #031L0211), the University Hospital Düsseldorf (Forschungskommission UKD), the United Mitochondrial Disease Foundation (UMDF), and the Deutsche Forschungsgemeinschaft (DFG) (PR1527/5-1). (#AZ.031A318 and #031L0211), the University Hospital Düsseldorf (Forschungskommission UKD), the United Mitochondrial Disease Foundation (UMDF), and the Deutsche Forschungsgemeinschaft (DFG) (PR1527/5-1).
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Inak, G., Henke, MT., Prigione, A. (2021). Bioenergetic Profiling of Human Pluripotent Stem Cells. In: Weissig, V., Edeas, M. (eds) Mitochondrial Medicine. Methods in Molecular Biology, vol 2277. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1270-5_24
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DOI: https://doi.org/10.1007/978-1-0716-1270-5_24
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-1269-9
Online ISBN: 978-1-0716-1270-5
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