Versatile open software to quantify cardiomyocyte and cardiac muscle contraction<i>in vitro</i>and<i>in vivo</i>

Sala, Luca; Meer, Berend J. van; Tertoolen, Leon G.J.; Bakkers, Jeroen; Bellin, Milena; Davis, Richard P.; Denning, Chris; Dieben, Michel A.E.; Eschenhagen, Thomas; Giacomelli, Elisa; Grandela, Catarina; Hansen, Arne; Holman, Eduard R.; Jongbloed, Monique R.M.; Kamel, Sarah M.; Koopman, Charlotte D.; Lachaud, Quentin; Mannhardt, Ingra; Mol, Mervyn P.H.; Orlova, Valeria V.; Passier, Robert; Ribeiro, Marcelo C.; Saleem, Umber; Smith, Godfrey L.; Mummery, Christine L.; Burton, Francis L.

doi:10.1101/160754

Cited by 106 publications

(164 citation statements)

References 27 publications

Supporting

Mentioning

154

Contrasting

Unclassified

Order By: Relevance

“…To date, several methods have been used to assess heartbeat in zebrafish hatchlings, medaka embryos, fish species, Drosophila, cardiomyocytes or mouse embryos 23,24,26,27,29,30,[39][40][41][42][43] . Some of these methods require transgenic embryos with fluorescent cardiomyocytes or erythrocytes to measure heartbeats 28,39 .…”

Section: Discussionmentioning

confidence: 99%

Automated high-throughput heart rate measurement in medaka and zebrafish embryos under physiological conditions

Gierten

Pylatiuk

Hammouda

et al. 2019

Preprint

View full text Add to dashboard Cite

Rationale: Accurate and efficient quantification of heartbeats in small fish models is an important readout to study cardiovascular biology, disease states and pharmacology at large scale. However, dependence on anesthesia, laborious sample orientation or requirement for fluorescent reporters have hampered the establishment of high-throughput heartbeat analysis.Objective: To overcome these limitations, we aimed to develop a high-throughput assay with automated heart rate scoring in medaka (Oryzias latipes) and zebrafish (Danio rerio) embryos under physiological conditions designed for genetic screens and drug discovery and validation. Methods and Results:We established an efficient screening assay employing automated label-free heart rate determination of randomly oriented, non-anesthetized specimen in microtiter plates. Automatically acquired bright-field data feeds into an easy-to-use HeartBeat software, a MATLAB algorithm with graphical user interface developed for automated quantification of heart rate and rhythm. Sensitivity of the assay and algorithm was demonstrated by profiling heart rates during entire embryonic development. Our analysis pipeline revealed acute temperature changes triggering rapid adaption of heart rates, which has implications for standardization of experimental layout. The approach is scalable and allows scoring of multiple embryos per well resulting in a throughput of >500 embryos per 96-well plate. In a proof of principle screen for compound testing, our assay captured concentration-dependent effects of nifedipine and terfenadine over time. Conclusion:A novel workflow and HeartBeat software provide efficient means for reliable and direct quantification of heart rate and rhythm of small fish in a physiological environment. Importantly, confounding factors such as anesthetics or laborious mounting are eliminated. We provide detailed profiles of embryonic heart rate dynamics in medaka and zebrafish as reference for future assay development. Ease of sample handling, automated imaging, physiological conditions and softwareassisted analysis now facilitate various large-scale applications ranging from phenotypic screening, interrogation of gene functions to cardiovascular drug development pipelines.

show abstract

Section: Discussionmentioning

confidence: 99%

Automated high-throughput heart rate measurement in medaka and zebrafish embryos under physiological conditions

Gierten

Pylatiuk

Hammouda

et al. 2019

Preprint

View full text Add to dashboard Cite

show abstract

“…(d) The low data output of the system offers a higher level of flexibility. The system does not require high‐performance computers or even an external server structure for data storage as required for video‐optical analysis techniques (e.g., Boudou et al, ; Hansen et al, ; Hayakawa et al, 2011; Ribeiro et al, ; Sala et al, ). This allows for continuous measurements up to days, ideal for the analysis of chronic toxicity (Jacob et al, ).…”

Section: Discussionmentioning

confidence: 99%

Piezo-bending actuators for isometric or auxotonic contraction analysis of engineered heart tissue

Mannhardt

Warncke

Trieu

et al. 2018

J Tissue Eng Regen Med

Self Cite

View full text Add to dashboard Cite

Engineered heart tissue (EHT) has proven as valuable tool for disease modelling, drug safety screening, and cardiac repair. Especially in combination with the stem cell technology, these in vitro models of the human heart have generated interest not only of basic cardiovascular researchers but also of regulatory authorities responsible for drug safety. A main limitation of 3D-based assays for evaluating cardiotoxicity is their limited throughput. We integrated piezo-bending actuators in a 24-well system for the generation of strip-like rat and human EHT attached to hollow, elastic silicone posts. Muscle contractions of EHTs induced a measurable electrical current in the piezo-bending actuators that could be analysed for contraction amplitude, frequency, and contraction and relaxation kinetics. Compared with the standard video-optical analysis of contractile activity, the new system allows for (a) the analysis of several tissues in parallel, (b) switching between auxotonic and isometric contractions by inserting a stiff metal post in the silicone post opposing the piezo actuator, (c) continuous measurement over days with low data volume (megabyte), (d) automated measurement without the necessity of adjustment of tissue position for video-optical analysis, (e) reduced complexity and costs, (f) high sensitivity of contraction detection, (g) calculation of absolute contraction force, and (h) suitability for variable tissue geometries. The new set-up for contraction analysis based on piezo-bending actuators is a promising new method for the parallel screening of EHT for pharmacological drug effects and other applications of muscle tissue engineering (e.g., skeletal muscle engineering or cardiac repair). KEYWORDS auxotonic, cardiac repair, contraction analysis, drug safety screening, engineered heart tissue, human induced pluripotent stem cells, isometric

show abstract

“…Microtissue contraction was recorded at 37°/5% CO2 in iCell maintenance media using a Nikon TE2000 (4x lens, 20 HZ). The software package musclemotion was used for all contraction analysis 64 .…”

Section: D Bioprinting Cardiac Microtissues and Contraction Imagingmentioning

confidence: 99%

3D bioprinting of high cell-density heterogeneous tissue models through spheroid fusion within self-healing hydrogels

Daly

Burdick

2020

Preprint

View full text Add to dashboard Cite

Cellular models are needed to study human development and disease in vitro, including the screening of drugs for toxicity and efficacy. However, current approaches are limited in the engineering of functional tissue models with requisite cell densities and heterogeneity to appropriately model cell and tissue behaviors. Here, we develop a new bioprinting approach to transfer spheroids into self-healing support hydrogels at high resolution, which enables their patterning and fusion into high-cell density microtissues of prescribed spatial organization. As an example application, we bioprint induced pluripotent stem cell-derived cardiac microtissue models with spatially controlled cardiomyocyte and fibroblast cell ratios to replicate the structural and functional features of scarred cardiac tissue that arise following myocardial infarction, including reduced contractility and irregular electrical activity. The bioprinted in vitro model is combined with functional readouts to probe how various pro-regenerative microRNA treatment regimes influence tissue regeneration and recovery of function as a result of cardiomyocyte proliferation. This method is useful for a range of biomedical applications, including the development of precision models to mimic diseases and for the screening of drugs, particularly where high cell densities and heterogeneity are important.

show abstract

Versatile open software to quantify cardiomyocyte and cardiac muscle contractionin vitroandin vivo

Abstract: Abstract

Cited by 106 publications

References 27 publications

Automated high-throughput heart rate measurement in medaka and zebrafish embryos under physiological conditions

Automated high-throughput heart rate measurement in medaka and zebrafish embryos under physiological conditions

Piezo-bending actuators for isometric or auxotonic contraction analysis of engineered heart tissue

3D bioprinting of high cell-density heterogeneous tissue models through spheroid fusion within self-healing hydrogels

Contact Info

Product

Resources

About