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Biology
实时测量人诱导多能干细胞来源心肌细胞中的钙和收缩力参数
实时测量人诱导多能干细胞来源心肌细胞中的钙和收缩力参数
JoVE Journal
Biology
This content is Free Access.
JoVE Journal Biology
Real-Time Measurements of Calcium and Contractility Parameters in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes

实时测量人诱导多能干细胞来源心肌细胞中的钙和收缩力参数

Full Text
3,594 Views
06:42 min
May 26, 2023

DOI: 10.3791/65326-v

Rafeeh Dinani1,2, Emmy Manders1,2,3, Michiel Helmes1,2,3, Lili Wang4, Bjorn C. Knollmann4, Diederik W. D. Kuster1,2, Jolanda van der Velden1,2

1Division of Physiology, Amsterdam UMC,Vrije University, 2Heart Failure & Arrhythmias,Amsterdam Cardiovascular Sciences, 3CytoCypher BV, 4Division of Clinical Pharmacology,Vanderbilt School of Medicine

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Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This study introduces a user-friendly method to measure the contractility and calcium handling of human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). The approach allows researchers to rapidly assess the effects of genetic mutations and pharmacological interventions, demonstrating its utility in drug screening and understanding cellular mechanisms.

Key Study Components

Research Area

  • Cardiomyocyte function
  • Drug interventions
  • Cellular pathophysiology

Background

  • hiPSC-CMs are valuable for studying cardiac diseases.
  • Understanding contraction and calcium dynamics is crucial for characterizing cardiomyocyte function.
  • A reproducible measurement platform is necessary for effective experimentation.

Methods Used

  • Optics-based measurement system for contractility and calcium measurements
  • Human-induced pluripotent stem cell-derived cardiomyocytes
  • Pixel correlation techniques and cytosolver analysis

Main Results

  • The method successfully captured synchronized contraction and relaxation along with calcium transients.
  • Application of beta-adrenergic receptor agonist iso increased beat frequency and improved contractile kinetics.
  • Data was efficiently analyzed using the Cytosolver program.

Conclusions

  • This method enhances the ability to study cardiac function at a cellular level.
  • It supports the development of therapeutic strategies through preclinical drug screening.

Frequently Asked Questions

What is the significance of using hiPSC-derived cardiomyocytes?
They provide a patient-specific model to study cardiac diseases and drug responses.
How does the measurement platform ensure reproducibility?
The platform standardizes conditions and measurement protocols, reducing variability.
What role does the calcium sensitive FloA-PhoR play in the experiment?
It allows for real-time measurements of intracellular calcium transients during contractions.
Can this method be applied to other cell types?
While designed for cardiomyocytes, the platform may be adaptable to other cell types with similar contractility features.
How does the platform contribute to drug screening?
It enables the rapid evaluation of drug effects on cardiac function, facilitating preclinical assessments.
What parameters are analyzed in the study of contractility?
Key parameters include resting frequency, time to peak contractility, and time to baseline contractility.
Is the method suitable for high-throughput screening?
Yes, the platform supports multiple plate formats for high-throughput applications.

本文描述了一种使用基于光学的平台在人诱导的多能干细胞衍生心肌细胞中进行收缩性和钙测量的既定方法。该平台使研究人员能够以快速且可重复的方式研究突变的影响以及对各种刺激的反应。

人类诱导的多能干细胞来源的心肌细胞是研究突变介导的心肌细胞功能变化以及定义应激源和药物干预效果的有力工具。需要一个平台,以用户友好和可重复的方式测量人诱导的多能干细胞来源心肌细胞的收缩力和钙处理。该方法使研究人员能够通过像素相关性研究人诱导的多能干细胞来源的心肌细胞的收缩性,并通过用钙敏感荧光团加载细胞同时测量细胞内钙瞬变。

通过使用该平台,可以在保存完好的温度环境和不同的板格式下进行配对测量,并立即访问数据。该方法能够研究细胞病理机制并评估化合物的作用和人iPSC来源的心肌细胞的功能,这可能有助于临床前药物筛选过程。

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