Abstract
Mitochondrial dynamics play an important role in numerous physiological and pathophysiological phenomena in the developing and adult human heart. Alterations in structural aspects of cellular mitochondrial composition as a function of changes in physiology can easily be visualized using fluorescence microscopy. Commonly, mitochondrial location, number, and morphology are reported qualitatively due to the lack of automated and user-friendly computer-based analysis tools. Mitochondrial Quantification using MATLAB (MQM) is a computer-based tool to quantitatively assess these parameters by analyzing fluorescently labeled mitochondria within the cell; in particular, MQM provides numerical information on the number, area, and location of mitochondria within a cell in a time-efficient, automated, and unbiased way. This chapter describes the use of MQM’s capabilities to quantify mitochondrial changes during human pluripotent stem cell (hPSC) differentiation into spontaneously contracting cardiomyocytes (SC-CMs), which follows physiological pathways of human heart development.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Park SJ, Shin JH, Jeong JI et al (2014) Down-regulation of mortalin exacerbates Abeta-mediated mitochondrial fragmentation and dysfunction. J Biol Chem 289:2195–2204
Rana A, Rera M, Walker DW (2013) Parkin overexpression during aging reduces proteotoxicity, alters mitochondrial dynamics, and extends lifespan. Proc Natl Acad Sci U S A 110:8638–8643
Abou-Sleiman PM, Muqit MM, Wood NW (2006) Expanding insights of mitochondrial dysfunction in Parkinson’s disease. Nat Rev Neurosci 7:207–219
Hashimoto M, Rockenstein E, Crews L et al (2003) Role of protein aggregation in mitochondrial dysfunction and neurodegeneration in Alzheimer’s and Parkinson’s diseases. Neuromolecular Med 4:21–36
Castellani R, Hirai K, Aliev G et al (2002) Role of mitochondrial dysfunction in Alzheimer’s disease. J Neurosci Res 70:357–360
Reddy PH, Beal MF (2008) Amyloid beta, mitochondrial dysfunction and synaptic damage: implications for cognitive decline in aging and Alzheimer’s disease. Trends Mol Med 14:45–53
Yoon Y, Galloway CA, Jhun BS et al (2011) Mitochondrial dynamics in diabetes. Antioxid Redox Signal 14:439–457
Lowell BB, Shulman GI (2005) Mitochondrial dysfunction and type 2 diabetes. Science 307:384–387
Desai SP, Bhatia SN, Toner M et al (2013) Mitochondrial localization and the persistent migration of epithelial cancer cells. Biophys J 104:2077–2088
Chen L, Knowlton AA (2011) Mitochondrial dynamics in heart failure. Congest Heart Fail 17:257–261
Ong SB, Hausenloy DJ (2010) Mitochondrial morphology and cardiovascular disease. Cardiovasc Res 88:16–29
Liu S, Bai Y, Huang J et al (2013) Do mitochondria contribute to left ventricular non-compaction cardiomyopathy? New findings from myocardium of patients with left ventricular non-compaction cardiomyopathy. Mol Genet Metab 109:100–106
Dhalla NS, Rangi S, Zieroth S et al (2012) Alterations in sarcoplasmic reticulum and mitochondrial functions in diabetic cardiomyopathy. Exp Clin Cardiol 17:115–120
Lopaschuk GD, Jaswal JS (2010) Energy metabolic phenotype of the cardiomyocyte during development, differentiation, and postnatal maturation. J Cardiovasc Pharmacol 56:130–140
Folmes CD, Dzeja PP, Nelson TJ et al (2012) Metabolic plasticity in stem cell homeostasis and differentiation. Cell Stem Cell 11:596–606
Mitra K, Lippincott-Schwartz J (2010) Analysis of mitochondrial dynamics and functions using imaging approaches. Curr Protoc Cell Biol Chapter 4:Unit 4.25.1–21
Chung S, Dzeja PP, Faustino RS et al (2007) Mitochondrial oxidative metabolism is required for the cardiac differentiation of stem cells. Nat Clin Pract Cardiovasc Med 4(Suppl 1):S60–S67
Hattori F, Chen H, Yamashita H et al (2010) Nongenetic method for purifying stem cell-derived cardiomyocytes. Nat Methods 7:61–66
Rafelski SM (2013) Mitochondrial network morphology: building an integrative, geometrical view. BMC Biol 11:71
Lian X, Zhang J, Azarin SM et al (2013) Directed cardiomyocyte differentiation from human pluripotent stem cells by modulating Wnt/beta-catenin signaling under fully defined conditions. Nat Protoc 8:162–175
Acknowledgment
This work was supported by AHA 13PRE1470078 (P.K.), NSF-CBET-1150854 (E.A.L.), NSF-EPS-1158862 (D.A.D.), and NSF-EEC-1063107 (K.M.D.). D.A.D. is now located at the Department of Biological Sciences, State University of New York at Oswego, Oswego, NY.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2015 Springer Science+Business Media New York
About this protocol
Cite this protocol
Kerscher, P., Bussie, B.S., DeSimone, K.M., Dunn, D.A., Lipke, E.A. (2015). Characterization of Mitochondrial Populations During Stem Cell Differentiation. In: Weissig, V., Edeas, M. (eds) Mitochondrial Medicine. Methods in Molecular Biology, vol 1264. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2257-4_37
Download citation
DOI: https://doi.org/10.1007/978-1-4939-2257-4_37
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2256-7
Online ISBN: 978-1-4939-2257-4
eBook Packages: Springer Protocols