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Related Experiment Video

Updated: Oct 18, 2025

Generating Self-Assembling Human Heart Organoids Derived from Pluripotent Stem Cells
08:56

Generating Self-Assembling Human Heart Organoids Derived from Pluripotent Stem Cells

Published on: September 15, 2021

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Generating Self-Assembling Human Heart Organoids Derived from Pluripotent Stem Cells.

Yonatan R Lewis-Israeli1, Brett D Volmert1, Mitchell A Gabalski1

  • 1Institute for Quantitative Health Science and Engineering, Division of Developmental and Stem Cell Biology, Michigan State University; Department of Biomedical Engineering, College of Engineering, Michigan State University.

Journal of Visualized Experiments : Jove
|October 4, 2021
PubMed
Summary

Researchers developed a new protocol to create complex human heart organoids (hHOs) from pluripotent stem cells. This advanced in vitro model mimics human cardiac development and disease, offering a powerful tool for scientific study.

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Area of Science:

  • Developmental Biology
  • Stem Cell Biology
  • Cardiovascular Research

Background:

  • Studying human cardiac development and disease in vitro is challenging due to the complexity of the human heart.
  • Organoids and organs-on-a-chip offer promising platforms for modeling complex in vivo phenotypes.
  • Efficient and reproducible in vitro models are crucial for advancing cardiovascular research.

Purpose of the Study:

  • To describe a protocol for generating highly complex human heart organoids (hHOs).
  • To utilize human pluripotent stem cells and stepwise developmental pathway activation for hHO generation.
  • To establish a robust platform for in vitro studies of human cardiac development and disease.

Main Methods:

  • Generation of embryoid bodies (EBs) in a 96-well ultra-low attachment plate format.
  • Three-step Wnt signaling modulation strategy (activation-inhibition-activation) to guide differentiation.
  • Immunofluorescence imaging and Fluo-4 live imaging for analysis of tissue complexity and function.

Main Results:

  • Successfully generated complex hHOs exhibiting first and second heart field specifications by day 11.
  • Day 15 hHOs displayed intricate tissues including myocardial, endocardial, epicardial layers, and internal chambers.
  • Organoids demonstrated robust beating and normal calcium activity, indicating functional cardiac tissue.

Conclusions:

  • The described protocol efficiently generates complex, self-organized human heart organoids (hHOs).
  • These hHOs serve as a valuable organ-like platform for in vitro studies of human cardiac development and disease.
  • The 96-well format ensures high efficiency, reproducibility, and scalability for generating cardiac organoids.