Contractility analysis of human engineered 3D heart tissues by an automatic tracking technique using a standalone application

PLoS One. 2022 Apr 14;17(4):e0266834. doi: 10.1371/journal.pone.0266834. eCollection 2022.

Abstract

The use of Engineered Heart Tissues (EHT) as in vitro model for disease modeling and drug screening has increased, as they provide important insight into the genetic mechanisms, cardiac toxicity or drug responses. Consequently, this has highlighted the need for a standardized, unbiased, robust and automatic way to analyze hallmark physiological features of EHTs. In this study we described and validated a standalone application to analyze physiological features of EHTs in an automatic, robust, and unbiased way, using low computational time. The standalone application "EHT Analysis" contains two analysis modes (automatic and manual) to analyzes the contractile properties and the contraction kinetics of EHTs from high speed bright field videos. As output data, the graphs of displacement, contraction force and contraction kinetics per file will be generated together with the raw data. Additionally, it also generates a summary file containing all the data from the analyzed files, which facilitates and speeds up the post analysis. From our study we highlight the importance of analyzing the axial stress which is the force per surface area (μN/mm2). This allows to have a readout overtime of tissue compaction, axial stress and leave the option to calculate at the end point of an experiment the physiological cross-section area (PSCA). We demonstrated the utility of this tool by analyzing contractile properties and compaction over time of EHTs made out of a double reporter human pluripotent stem cell (hPSC) line (NKX2.5EGFP/+-COUP-TFIImCherry/+) and different ratios of human adult cardiac fibroblasts (HCF). Our standalone application "EHT Analysis" can be applied for different studies where the physiological features of EHTs needs to be analyzed under the effect of a drug compound or in a disease model.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • Drug Evaluation, Preclinical
  • Heart / physiology
  • Humans
  • Myocardial Contraction*
  • Myocytes, Cardiac
  • Tissue Engineering* / methods

Associated data

  • figshare/10.6084/m9.figshare.19492385
  • figshare/10.6084/m9.figshare.19492211
  • figshare/10.6084/m9.figshare.19492229

Grants and funding

This work was supported by the Netherlands Organ-on-Chip Initiative, an NWO Gravitation project (024.003.001) funded by the Ministry of Education, Culture and Science of the government of the Netherlands; grant MONACO-SPRINT (LSHM20063) co-funded by the PPP Allowance made available by Health-Holland; Top Sector Life Sciences & Health and Health Holland Organ On Chip grant. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.