title: Automated Video-Based Analysis of Contractility and Calcium Flux in Human-Induced Pluripotent Stem Cell-Derived Cardiomyocytes Cultured over Different Spatial Scales.
abstract: Contractile motion is the simplest metric of cardiomyocyte health in vitro, but unbiased quantification is challenging. We describe a rapid automated method, requiring only standard video microscopy, to analyze the contractility of human-induced pluripotent stem cell-derived cardiomyocytes (iPS-CM). New algorithms for generating and filtering motion vectors combined with a newly developed isogenic iPSC line harboring genetically encoded calcium indicator, GCaMP6f, allow simultaneous user-independent measurement and analysis of the coupling between calcium flux and contractility. The relative performance of these algorithms, in terms of improving signal to noise, was tested. Applying these algorithms allowed analysis of contractility in iPS-CM cultured over multiple spatial scales from single cells to three-dimensional constructs. This open source software was validated with analysis of isoproterenol response in these cells, and can be applied in future studies comparing the drug responsiveness of iPS-CM cultured in different microenvironments in the context of tissue engineering.
authors.affiliation: 1 Gladstone Institute of Cardiovascular Disease , San Francisco, California.
authors.forename: Nathaniel
authors.initials: N
authors.lastname: Huebsch
authors.forename: Peter
authors.initials: P
authors.lastname: Loskill
authors.forename: Mohammad A
authors.initials: MA
authors.lastname: Mandegar
authors.forename: Natalie C
authors.initials: NC
authors.lastname: Marks
authors.forename: Alice S
authors.initials: AS
authors.lastname: Sheehan
authors.forename: Zhen
authors.initials: Z
authors.lastname: Ma
authors.forename: Anurag
authors.initials: A
authors.lastname: Mathur
authors.forename: Trieu N
authors.initials: TN
authors.lastname: Nguyen
authors.forename: Jennie C
authors.initials: JC
authors.lastname: Yoo
authors.forename: Luke M
authors.initials: LM
authors.lastname: Judge
authors.forename: C Ian
authors.initials: CI
authors.lastname: Spencer
authors.forename: Anand C
authors.initials: AC
authors.lastname: Chukka
authors.forename: Caitlin R
authors.initials: CR
authors.lastname: Russell
authors.forename: Po-Lin
authors.initials: PL
authors.lastname: So
authors.forename: Bruce R
authors.initials: BR
authors.lastname: Conklin
authors.forename: Kevin E
authors.initials: KE
authors.lastname: Healy
mesh_terms: D000465:Algorithms; D002118:Calcium / Q000378:metabolism*; D002454:Cell Differentiation; D002478:Cells, Cultured / Q000166:cytology; D006801:Humans; D007091:Image Processing, Computer-Assisted; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D018715:Microscopy, Video / Q000379:methods*; D009200:Myocardial Contraction; D032383:Myocytes, Cardiac / Q000166:cytology*; D018408:Patch-Clamp Techniques; D010363:Pattern Recognition, Automated; D015398:Signal Transduction; D059629:Signal-To-Noise Ratio; D012984:Software
chemical_list: D002118:Calcium