Analysis code for "Hypertrophy changes 3D shape of hiPSC-cardiomyocytes: Implications for cellular maturation in regenerative medicine"

Description

Abstract:
Advances in the use of human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes for heart regeneration and in vitro disease models demand a greater understanding of how these cells grow and mature in 3-dimensional space. In this study, we developed an analysis methodology of single cardiomyocytes plated on 2D surfaces to assess their 3D myofilament volume and its z-height distribution, or shape, upon hypertrophic stimulation via phenylephrine (PE) treatment or long-term culture (“aging”). Cardiomyocytes were fixed and labeled with α-actinin for confocal microscopy imaging to obtain z-stacks for 3D myofilament volume analysis. In primary neonatal rat ventricular myocytes (NRVMs), area increased 72% with PE, while volume increased 31%. In hiPSC-cardiomyocytes, area increased 70% with PE and 4-fold with aging; however, volume increased significantly only with aging by 2.3-fold. Analysis of z-height myofilament volume distribution in hiPSC-cardiomyocytes revealed a shift from a fairly uniform distribution in control cells to a basally located volume in a more flat and spread morphology with PE and even more so with aging, a shape that was akin to all NRVMs analyzed. These results suggest that 2D area is not a sufficient measure of hiPSC-cardiomyocyte growth and maturation, and that changes in 3D volume and its distribution are essential for understanding hiPSC-cardiomyocyte biology for disease modeling and regenerative medicine applications.
Notes:
This research was supported by funding from the National Institutes of Health, National Heart, Lung and Blood Institute, grant ID HL115123

Access Conditions

Use and Reproduction
Available under a GNU General Public License

Citation

Rupert, Cassady, Chang, Heidi, and Coulombe, Kareen, "Analysis code for 'Hypertrophy changes 3D shape of hiPSC-cardiomyocytes: Implications for cellular maturation in regenerative medicine'" (2016). Brown University Open Data Collection. Brown Digital Repository. Brown University Library. https://doi.org/10.7301/Z0WS8R5F

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  • CellProfiler analysis code from "Hypertrophy changes 3D shape of hiPSC-cardiomyocytes: Implications for cellular maturation in regenerative medicine"
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  • MATLAB analysis code from "Hypertrophy changes 3D shape of hiPSC-cardiomyocytes: Implications for cellular maturation in regenerative medicine"
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