Characterization of mitochondrial populations during stem cell differentiation

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 fl uorescence microscopy. Commonly, mitochondrial location, number, and morphology are reported qualitatively due to the lack of automated and user-friendly computer-based analysis tools. Mitochondrial Quantifi cation using MATLAB (MQM) is a computer-based tool to quantitatively assess these parameters by analyzing fl uorescently 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-effi cient, 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.