Morphometric Analysis of Human Embryonic Stem Cell-Derived Ventricular Cardiomyocytes: Determining the Maturation State of a Population by Quantifying Parameters in Individual Cells

Figure 3

Effect of isoproterenol and oleic acid on the orientation of myofibrils in the hESC-vCMs. ((a) panel (A)–(a) panel (C)) The orientation of the myofibrils localized within hESC-vCMs was determined by calculating the slope of lines placed along the length of the myofibrils. ((a) panel (A)) hESC-vCMs with the myofibrils arranged in parallel exhibited a low slope variance (SV) value (e.g., 0.02 AU), when compared with ((a) panel (B)) cells where the myofibrils were arranged in a concentric manner and exhibit an SV of ~124 AU or ((a) panel (C)) where the myofibrils were arranged in an irregular orientation when the SV was greater again (e.g., ~480 AU). Scale bar is 30 μm. (b–g) Histograms to show the distribution of cells (in %) exhibiting a particular variance value in (b) untreated controls and (c–g) following treatment with (c) 0.3 μM isoproterenol, (d) 100 μM oleic acid, (e) 200 μM oleic acid, (f) 0.3 μM isoproterenol plus 100 μM oleic acid, or (g) 0.3 μM isoproterenol plus 200 μM oleic acid. for each treatment group and the untreated controls. The grey bar and grey dashed lines indicate the percentage of cells in the untreated control group with the lowest myofibrillar SV value.