Optical-flow based non-invasive analysis of cardiomyocyte contractility

Czirók, András; Isai, Dona Greta; Kosa, Edina; Rajasingh, Sheeja; Kinsey, William H.; Neufeld, Zoltán; Rajasingh, Johnson

doi:10.1038/s41598-017-10094-7

Cited by 50 publications

(40 citation statements)

References 40 publications

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“…SV1), as well as increases in other parameters of sheet contractility (for extended data see Supporting Information Fig. S3) [44,45]. Although some of these methods may be sensitive to the rate of beating, we found no significant difference between the spontaneous beat frequency in hPSC-CMs from pIC-treated and -untreated CPCs (Fig.…”

Section: Results Pic Priming Of Cpcs Accelerates Hpsc-cm Maturationmentioning

confidence: 69%

Epigenetic Priming of Human Pluripotent Stem Cell-Derived Cardiac Progenitor Cells Accelerates Cardiomyocyte Maturation

et al. 2019

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Human pluripotent stem cell‐derived cardiomyocytes (hPSC‐CMs) exhibit a fetal phenotype that limits in vitro and therapeutic applications. Strategies to promote cardiomyocyte maturation have focused interventions on differentiated hPSC‐CMs, but this study tests priming of early cardiac progenitor cells (CPCs) with polyinosinic‐polycytidylic acid (pIC) to accelerate cardiomyocyte maturation. CPCs were differentiated from hPSCs using a monolayer differentiation protocol with defined small molecule Wnt temporal modulation, and pIC was added during the formation of early CPCs. pIC priming did not alter the expression of cell surface markers for CPCs (>80% KDR+/PDGFRα+), expression of common cardiac transcription factors, or final purity of differentiated hPSC‐CMs (∼90%). However, CPC differentiation in basal medium revealed that pIC priming resulted in hPSC‐CMs with enhanced maturity manifested by increased cell size, greater contractility, faster electrical upstrokes, increased oxidative metabolism, and more mature sarcomeric structure and composition. To investigate the mechanisms of CPC priming, RNAseq revealed that cardiac progenitor‐stage pIC modulated early Notch signaling and cardiomyogenic transcriptional programs. Chromatin immunoprecipitation of CPCs showed that pIC treatment increased deposition of the H3K9ac activating epigenetic mark at core promoters of cardiac myofilament genes and the Notch ligand, JAG1. Inhibition of Notch signaling blocked the effects of pIC on differentiation and cardiomyocyte maturation. Furthermore, primed CPCs showed more robust formation of hPSC‐CMs grafts when transplanted to the NSGW mouse kidney capsule. Overall, epigenetic modulation of CPCs with pIC accelerates cardiomyocyte maturation enabling basic research applications and potential therapeutic uses. Stem Cells 2019;37:910–923

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Section: Results Pic Priming Of Cpcs Accelerates Hpsc-cm Maturationmentioning

confidence: 69%

Epigenetic Priming of Human Pluripotent Stem Cell-Derived Cardiac Progenitor Cells Accelerates Cardiomyocyte Maturation

et al. 2019

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“…The optical flow-based method does not distinguish between active and passive (elastic response of the adjacent cell layer) contractility. To identify contractile centers, we estimated the convergence maps of the displacement field as its negative divergence from optical flow data d(t,x) as previously described (Czirok et al, 2017).…”

Section: Convergence Analysismentioning

confidence: 99%

Scalable Biomimetic Coaxial Aligned Nanofiber Cardiac Patch: A Potential Model for “Clinical Trials in a Dish”

Kumar

Sridharan

Palaniappan

et al. 2020

Front. Bioeng. Biotechnol.

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Recent advances in cardiac tissue engineering have shown that human inducedpluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) cultured in a threedimensional (3D) micro-environment exhibit superior physiological characteristics compared with their two-dimensional (2D) counterparts. These 3D cultured hiPSC-CMs have been used for drug testing as well as cardiac repair applications. However, the fabrication of a cardiac scaffold with optimal biomechanical properties and high biocompatibility remains a challenge. In our study, we fabricated an aligned polycaprolactone (PCL)-Gelatin coaxial nanofiber patch using electrospinning. The structural, chemical, and mechanical properties of the patch were assessed by scanning electron microscopy (SEM), immunocytochemistry (ICC), Fourier-transform infrared spectroscopy (FTIR)-spectroscopy, and tensile testing. hiPSC-CMs were cultured on the aligned coaxial patch for 2 weeks and their viability [lactate dehydrogenase (LDH assay)], morphology (SEM, ICC), and functionality [calcium cycling, multielectrode array (MEA)] were assessed. Furthermore, particle image velocimetry (PIV) and MEA were used to evaluate the cardiotoxicity and physiological functionality of the cells in response to cardiac drugs. Nanofibers patches were comprised of highly aligned core-shell fibers with an average diameter of 578 ± 184 nm. Acellular coaxial patches were significantly stiffer than gelatin alone with an ultimate tensile strength of 0.780 ± 0.098 MPa, but exhibited gelatin-like biocompatibility. Furthermore, hiPSC-CMs cultured on the surface of these aligned coaxial patches (3D cultures) were elongated and rod-shaped with well-organized sarcomeres, as observed by the expression of cardiac troponin-T and α-sarcomeric actinin. Additionally, hiPSC-CMs cultured on these coaxial patches formed a functional syncytium evidenced by the expression of connexin-43 (Cx-43) and

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“…To characterize tissue deformation, we first apply our non-invasive, optical flowbased method described in (Czirok et al, 2017) for each image of the recording.…”

Section: Optical Flow-based Analysis Of Local Tissue Rotationmentioning

confidence: 99%

“…To quantify the tissue deformation induced by the ferromagnetic aggregates, we modified our image analysis tools used to characterize cardiomyocyte beating activity (Czirok et al, 2017). We compared a sequence of images to a common reference frame by PIV analysis, yielding a displacement field (u).…”

Section: Tissue Deformation Forced By External Fieldsmentioning

confidence: 99%

Viscoelastic properties of ECM-rich embryonic microenvironments

Ákos

Isai

Rajasingh

et al. 2019

Preprint

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The material properties of tissues and their mechanical state is an important factor during development, disease, regenerative medicine and tissue engineering. Here we describe a microrheological measurement technique utilizing aggregates of microinjected ferromagnetic nickel particles to probe the viscoelastic properties of embryonic tissues. Quail embryos were cultured in a plastic incubator chamber located at the center of two pairs of crossed electromagnets. We estimate the Young's modulus of the ECM-rich region separating the mesoderm and endoderm in HamburgerHamilton stage 6-10 quail embryos as 300±100 Pa. We found a pronounced viscoelastic behavior consistent with a Zener (standard generalized solid) model. The viscoelastic response is about 45% of the total response, with a characteristic relaxation time of 1.3 sec.

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Optical-flow based non-invasive analysis of cardiomyocyte contractility

Cited by 50 publications

References 40 publications

Epigenetic Priming of Human Pluripotent Stem Cell-Derived Cardiac Progenitor Cells Accelerates Cardiomyocyte Maturation

Epigenetic Priming of Human Pluripotent Stem Cell-Derived Cardiac Progenitor Cells Accelerates Cardiomyocyte Maturation

Scalable Biomimetic Coaxial Aligned Nanofiber Cardiac Patch: A Potential Model for “Clinical Trials in a Dish”

Viscoelastic properties of ECM-rich embryonic microenvironments

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