Rianú: Multi-Tissue Tracking Software for Increased Throughput of Engineered Cardiac Tissue Screening
13 Pages Posted: 1 Mar 2022 Last revised: 8 Mar 2022
Background and Objective: The field of cardiac tissue engineering has provided valuable three-dimensional species-specific models of the human myocardium in the form of human Engineered Cardiac Tissues (hECTs) and similar constructs. However, hECT systems are bottlenecked by software that can only collect data from one tissue at a time, even in multi-tissue bioreactors, which complicates experimental protocols and limits throughput in phenotypic and therapeutic screening applications. In this study, we aimed to develop a software to overcome the limitations of current systems and provide a tool that will increase throughput during analysis in cardiac tissue engineering.
Methods: To achieve this, we developed Rianú, an open-source web application capable of simultaneously tracking multiple hECTs on flexible end-posts. This software is operating system agnostic and deployable on a remote server, accessible via a web browser with no local hardware or software requirements. The software incorporates object-tracking capabilities for multiple objects simultaneously, an algorithm for twitch tracing analysis and contractile force calculation, and a data compilation system for comparative analysis within and amongst groups. Validation tests were performed using in-silico and in-vitro experiments for comparison with established methods and interventions. Statistical analysis was performed using linear least-squares regression, Student’s t-test or One way ANOVA.
Results: Rianú was able to detect the displacement of the flexible end-posts with a sub-pixel sensitivity of 0.555 px/post (minimum increment in post displacement) and a lower limit of 1.665 px/post (minimum post displacement). Compared to our gold standard for contractility assessment, Rianú had a high correlation for all parameters analyzed (ranging from R2 = 0.7514 to R2 = 0.9695), demonstrating its high accuracy and reliability.
Conclusions: Rianú provides simultaneous tracking of multiple hECTs, expediting the recording and analysis processes, and simplifies time-based intervention studies. It also allows data collection from different formats and has scale-up capabilities proportional to the number of tissues per field of view. These capabilities will enhance throughput of hECTs and similar assays for in vitro analysis in disease modeling and drug screening applications.
Funding Information: This work was funded by the National Heart, Lung, and Blood Institute of the National Institutes of Health under Award Number R03HL154286 (ICT), K01HL133424 (ICT) and R01HL132226 (KDC).
Declaration of Interests: K.D.C. holds equity in NovoHeart Holdings. NovoHeart did not contribute to the funding, planning, or execution of this study; however, the study outcomes could potentially have a financial impact on NovoHeart. The other authors declare that they have no competing interests.
Keywords: tissue engineering, object tracking, cardiac muscle function, human engineered cardiac tissues, contractility analysis, drug screening
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