High content analysis identifies unique morphological features of reprogrammed cardiomyocytes

Matthew D. Sutcliffe, Philip M. Tan, Antonio Fernandez-Perez, Young Jae Nam, Nikhil V. Munshi, Jeffrey J. Saucerman

Research output: Contribution to journalArticle

Abstract

Direct reprogramming of fibroblasts into cardiomyocytes is a promising approach for cardiac regeneration but still faces challenges in efficiently generating mature cardiomyocytes. Systematic optimization of reprogramming protocols requires scalable, objective methods to assess cellular phenotype beyond what is captured by transcriptional signatures alone. To address this question, we automatically segmented reprogrammed cardiomyocytes from immunofluorescence images and analyzed cell morphology. We also introduce a method to quantify sarcomere structure using Haralick texture features, called SarcOmere Texture Analysis (SOTA). We show that induced cardiac-like myocytes (iCLMs) are highly variable in expression of cardiomyocyte markers, producing subtypes that are not typically seen in vivo. Compared to neonatal mouse cardiomyocytes, iCLMs have more variable cell size and shape, have less organized sarcomere structure, and demonstrate reduced sarcomere length. Taken together, these results indicate that traditional methods of assessing cardiomyocyte reprogramming by quantifying induction of cardiomyocyte marker proteins may not be sufficient to predict functionality. The automated image analysis methods described in this study may enable more systematic approaches for improving reprogramming techniques above and beyond existing algorithms that rely heavily on transcriptome profiling.

Original languageEnglish (US)
Article number1258
JournalScientific Reports
Volume8
Issue number1
DOIs
StatePublished - Dec 1 2018

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Cardiac Myocytes
Sarcomeres
Cell Shape
Gene Expression Profiling
Cell Size
Fluorescent Antibody Technique
Regeneration
Fibroblasts
Phenotype

ASJC Scopus subject areas

  • General

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High content analysis identifies unique morphological features of reprogrammed cardiomyocytes. / Sutcliffe, Matthew D.; Tan, Philip M.; Fernandez-Perez, Antonio; Nam, Young Jae; Munshi, Nikhil V.; Saucerman, Jeffrey J.

In: Scientific Reports, Vol. 8, No. 1, 1258, 01.12.2018.

Research output: Contribution to journalArticle

Sutcliffe, Matthew D. ; Tan, Philip M. ; Fernandez-Perez, Antonio ; Nam, Young Jae ; Munshi, Nikhil V. ; Saucerman, Jeffrey J. / High content analysis identifies unique morphological features of reprogrammed cardiomyocytes. In: Scientific Reports. 2018 ; Vol. 8, No. 1.
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