Automated analysis of human cardiomyocytes dynamics with holographic image-based tracking for cardiotoxicity
Ezat Ahamadzadeh1, Keyvan Jaferzadeh2, Seonghwan Park1
1Department of Robotics Engineering, Daegu Gyeongbuk Institute of Science & Technology (DGIST), Hyeonpung-eup, Dalseong-gun, Daegu, 42988, South Korea.
Insights
A new holographic imaging platform non-invasively tracks human-induced pluripotent stem cell-derived cardiomyocytes (hiPS-CMs) for automated cardiotoxicity screening. This method quantifies single-cell dynamics, revealing drug effects on heart cell beating patterns.
Area of Science:
- Biomedical Engineering
- Stem Cell Biology
- Cardiovascular Research
Background:
- Cardiotoxicity screening is crucial for drug development.
- Existing methods for analyzing cardiomyocyte (CM) mechanical activity can be costly and complex.
- Human-induced pluripotent stem cell-derived cardiomyocytes (hiPS-CMs) offer a promising model for cardiac studies.
Purpose of the Study:
- To develop and validate a novel, non-invasive, automated platform for quantitative single-cell analysis of hiPS-CMs.
- To utilize holographic imaging and optical flow for precise characterization of CM contractile dynamics.
- To establish a cost-effective method for cardiotoxicity screening using hiPS-CMs.
Main Methods:
- Combined dense Farneback optical flow with holographic imaging informatics for CM motion analysis.
- Quantitatively characterized single-cell motion, synchronization, and speed.
- Assessed platform reliability through motion characterization, synchronization, fixed vs. live CM speed, and noise sensitivity.
- Evaluated drug effects using isoprenaline and E-4031 on hiPS-CMs.
Main Results:
- The platform accurately characterized hiPS-CM contractile motion and kinetics at the single-cell level.
- Isoprenaline increased beat frequency by enhancing action potential (AP) speed and shortening the resting period.
- E-4031 decreased beat frequency by reducing AP speed and prolonging the resting period.
- Demonstrated the platform's ability to detect pharmacological effects of cardiovascular drugs.
Conclusions:
- The developed holographic imaging platform provides a reliable and automated method for analyzing hiPS-CM dynamics.
- This approach enables quantitative assessment of cardiotoxicity by characterizing single-cell mechanical responses to drugs.
- Offers valuable insights into hiPS-CM kinetics for improved drug safety evaluation.
Abstract:
This paper proposes a new non-invasive, low-cost, and fully automated platform to quantitatively analyze dynamics of human-induced pluripotent stem cell-derived cardiomyocytes (hiPS-CMs) at the single-cell level by holographic image-based tracking for cardiotoxicity screening. A dense Farneback optical flow method and holographic imaging informatics were combined to characterize the contractile motion of a single CM, which obviates the need for costly equipment to monitor a CM's mechanical beat activity. The reliability of the proposed platform was tested by single-cell motion characterization, synchronization analysis, motion speed measurement of fixed CMs versus live CMs, and noise sensitivity. The applicability of the motion characterization method was tested to determine the pharmacological effects of two cardiovascular drugs, isoprenaline (166 nM) and E-4031 (500 μM). The experiments were done using single CMs and multiple cells, and the results were compared to control conditions. Cardiomyocytes responded to isoprenaline by increasing the action potential (AP) speed and shortening the resting period, thus increasing the beat frequency. In the presence of E-4031, the AP speed was decreased, and the resting period was prolonged, thus decreasing the beat frequency. The findings offer insights into single hiPS-CMs' contractile motion and a deep understanding of their kinetics at the single-cell level for cardiotoxicity screening.
