Rianú:多组织跟踪软件,用于增加工程心脏组织筛选的吞吐量

Jack F. Murphy, Kevin D. Costa, Irene C. Turnbull
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引用次数: 0

摘要

背景:组织工程领域已经以人类工程心脏组织(hECTs)和类似结构的形式提供了有价值的人类心肌的三维物种特异性模型。然而,hECT系统经常因缺乏能够一次从多个组织收集数据的公开可用软件而受到瓶颈,即使在多组织生物反应器中也是如此,这限制了表型和治疗筛选应用的吞吐量。方法:我们开发了Rianú,这是一个开源的网络应用程序,能够同时跟踪灵活终端上的多个hECTs。该软件与操作系统无关,可部署在远程服务器上,可通过web浏览器访问,无需本地硬件或软件。该软件结合了同时对多个对象的对象跟踪功能、用于抽搐跟踪分析和收缩力计算的算法,以及用于组内和组间比较分析的数据汇编系统。使用计算机和体外实验进行验证测试,以与既定方法和干预措施进行比较。结果:Rianú能够检测柔性端柱的位移,亚像素灵敏度为0.555像素/柱(柱位移的最小增量),下限为1.665像素/柱。与我们建立的收缩力评估参考相比,Rianú对所有分析参数都有很高的相关性(范围从R2=0.7514到R2=0.9695),证明了其高准确性和可靠性。结论:Rianú可以同时跟踪多个hECTs,加快记录和分析过程,并简化基于时间的干预研究。它还允许从不同格式收集数据,并具有与每个视场的组织数量成比例的放大功能。这些能力将提高hECTs和类似测定的产量,用于疾病建模和药物筛选应用中的体外分析。
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Rianú: Multi-tissue tracking software for increased throughput of engineered cardiac tissue screening

Background:

The field of 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 often bottlenecked by a lack of openly available software that can collect data from multiple tissues at a time, even in multi-tissue bioreactors, which limits throughput in phenotypic and therapeutic screening applications.

Methods:

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.

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 established reference 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.

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