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

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Title
Analysis code for "Hypertrophy changes 3D shape of hiPSC-cardiomyocytes: Implications for cellular maturation in regenerative medicine"
Contributors
Rupert, Cassady (author)
Chang, Heidi (author)
Coulombe, Kareen (author)
Doi
10.7301/Z0WS8R5F
Date Created
2016-02-26
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.
Keywords
Confocal microscopy
Image analysis
Phenyephrine
Aging
Myofilament volume
Notes
This research was supported by funding from the National Institutes of Health, National Heart, Lung and Blood Institute, grant ID HL115123
Extent
MATLAB script, CellProfiler pipeline
Access Conditions
Available under a GNU General Public License
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). Data for Publications. Brown Digital Repository. Brown University Library. https://doi.org/10.7301/Z0WS8R5F

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