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

Full Metadata

Overview

Year:
2016
Contributor:
Rupert, Cassady (author)
Chang, Heidi (author)
Coulombe, Kareen (author)
Subject:
Confocal microscopy
Image analysis
Phenyephrine
Aging
Myofilament volume
Extent:
MATLAB script, CellProfiler pipeline
DOI
https://doi.org/10.7301/Z0WS8R5F

Files

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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Collection:

  • Brown University Open … Thumbnail

    Brown University Open Data Collection

    This collection contains open and publicly-funded data sets created by Brown University faculty and student researchers. Increasingly, publishers, and funders are requiring that protocols, data sets, metadata, and code underlying published research be retained and preserved, their locations cited within …
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