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Bioengineering
使用成熟的人诱导多能干细胞来源心肌细胞单层进行高通量心脏毒性筛选
使用成熟的人诱导多能干细胞来源心肌细胞单层进行高通量心脏毒性筛选
JoVE Journal
Bioengineering
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JoVE Journal Bioengineering
High-Throughput Cardiotoxicity Screening Using Mature Human Induced Pluripotent Stem Cell-Derived Cardiomyocyte Monolayers

使用成熟的人诱导多能干细胞来源心肌细胞单层进行高通量心脏毒性筛选

Full Text
2,535 Views
14:03 min
March 24, 2023

DOI: 10.3791/64364-v

Andre Monteiro da Rocha1, Andrew Allan2, Travis Block3, Jeffery Creech1, Todd J. Herron1

1Frankel Cardiovascular Regeneration Core Laboratory, Cardiovascular Medicine-Internal Medicine,University of Michigan, Ann Arbor, 2CAIRN Research, 3StemBioSys, Inc.

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Overview

This protocol enables the maturation of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) to an adult-like phenotype, enhancing their predictive value in cardiotoxicity screening. The method is designed for high throughput, allowing for efficient optical mapping of calcium or voltage changes.

Key Study Components

Area of Science

  • Cardiovascular research
  • Stem cell biology
  • Toxicology

Background

  • hiPSC-CMs are a promising alternative to animal models for cardiotoxicity testing.
  • The immature phenotype of hiPSC-CMs limits their predictive capabilities.
  • Protocols for maturation are essential for advancing their use in research.
  • This study presents a method to achieve rapid maturation of hiPSC-CMs.

Purpose of Study

  • To develop a protocol for maturing hiPSC-CMs to an adult-like phenotype.
  • To improve the predictive value of hiPSC-CMs in cardiotoxicity screening.
  • To facilitate high throughput analysis of cardiac function.

Methods Used

  • Preparation of maturation-inducing extracellular matrix (MECM) plates.
  • Washing MECM plates with Hank's balanced salt solution (HBSS).
  • Plating hiPSC-CMs on the prepared MECM plates.
  • Optical mapping of calcium or voltage changes in matured hiPSC-CMs.

Main Results

  • The protocol successfully matures hiPSC-CMs to an adult-like phenotype.
  • Mature hiPSC-CMs demonstrate improved predictive capabilities for cardiotoxicity.
  • The method supports high throughput screening for drug testing.
  • Optical mapping reveals significant calcium and voltage changes in matured cells.

Conclusions

  • This maturation protocol enhances the utility of hiPSC-CMs in cardiotoxicity research.
  • High throughput capabilities make it suitable for large-scale studies.
  • The findings support the potential of hiPSC-CMs as a reliable model for cardiac studies.

Frequently Asked Questions

What are hiPSC-CMs?
Human induced pluripotent stem cell-derived cardiomyocytes are heart cells derived from stem cells that can be used for research.
Why is maturation of hiPSC-CMs important?
Maturation improves their resemblance to adult heart cells, enhancing their predictive value in cardiotoxicity screening.
What is the main advantage of this protocol?
It allows for the rapid maturation of hiPSC-CMs in a high throughput format, facilitating efficient testing.
How does optical mapping work?
Optical mapping involves using fluorescent indicators to visualize changes in calcium or voltage in cardiac cells.
Can this protocol be used for drug screening?
Yes, it can be used to screen drugs for cardiotoxicity by assessing the function of matured hiPSC-CMs.
Who demonstrated the procedure?
The procedure was demonstrated by Jeffrey Creech, a research associate from the laboratory.

人诱导多能干细胞来源的心肌细胞(hiPSC-CMs)为使用动物进行临床前心脏毒性筛查提供了一种替代方案。hiPSC-CMs在临床前毒性筛查中广泛采用的一个限制是细胞的不成熟,胎儿样表型。这里介绍的是用于hiPSC-CMs稳健和快速成熟的方案。

该协议意义重大,因为它使研究人员能够将商业或内部制造的hiPSC-CMs成熟为成人样表型,从而显着提高hiPSC-CMs在心脏毒性筛查中的预测价值。该协议的主要优点是它以高通量形式提供成熟的hiPSC-CM,用于钙或电压变化的光学映射。该方案可用于研究疾病机制或筛查药物的心脏毒性。

演示该程序的将是我实验室的研究助理Jeffrey Creech。首先在心肌细胞铺板前 1 小时用 200 微升汉克平衡盐溶液或 HBSS 洗涤成熟诱导细胞外基质或 MECM 板两次。保持井水充足。

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