How to Prepare Heart Organoids

How to Prepare Heart Organoids

The heart is a muscular organ in vertebrates that pumps blood throughout the circulatory system, providing the necessary force for blood flow. Due to its complex structure, the heart is one of the most challenging organs to model in vitro.

Organoids are 3D cell structures cultured in vitro that mimic the architecture and function of organs in vivo, allowing for more accurate preservation of the biological characteristics and functionality of cells in the body. In recent years, the research team led by Mendjan S has induced stem cells to self-organize by activating all six known signaling pathways involved in embryonic heart development in a specific sequence, creating the world's first in vitro self-organizing heart organoid model. This breakthrough opens new directions and approaches for studying cardiovascular diseases.

Figure 1. Establishment of Heart Organoids from Human Pluripotent Stem Cells (PMID: 34019794)

Cultivation Plan for Heart Organoids

Based on the article published by Mendjan S[1], we have compiled a cultivation plan for generating Heart Organoids from human pluripotent stem cells (hPSC). This model can spontaneously form cavities, autonomously beat without the need for scaffolding support, and can mobilize cardiac fibroblasts to migrate and repair injuries after damage.


Figure 2. Process of hPSC Differentiation into Heart Organoids (PMID: 34019794) 

1. Mesoderm Induction

   hPSCs (WTC cells) are seeded onto ultra-low attachment 96-well plates and cultured in FLyAB(Ins) medium (CDM medium containing 30 ng/mL FGF2, 5 μM LY294002, 50 ng/mL Activin A, 10 ng/mL BMP4, 1 μg/mL insulin, 4-8 μM CHIR99021, 5 μM Y-27632) for 36-40 hours.

2. Cardiac Mesoderm

   Medium is changed to BFIIWPRa medium (CDM medium containing 10 ng/mL BMP4, 8 ng/mL FGF2, 10 μg/mL insulin, 5 μM IWP2, 0.5 μM retinoic acid, 200 ng/mL VEGF-A) and cultured for 4 days with daily medium changes.

3. Specific CM Differentiation

   Medium is changed to BFI medium (containing 10 ng/mL BMP4, 8 ng/mL FGF2, 10 μg/mL insulin, 100 ng/mL VEGF-A) and cultured for 2 days with daily medium changes. 

Figure 3. Changes in Cell Cluster Size and Bright-Field Images at Various Time Points during hPSC Differentiation into Heart Organoids (PMID: 34019794)

4. Organoid Cultivation

   Medium is changed to CDM medium (containing 100 ng/mL VEGF-A) and cultivation is continued, with medium changes every two days.  

Related products

Product Type

Product Name

Catalog Number


Recombinant protein

Recombinant Human/Mouse/Rat Activin A Protein


10 μg/100 μg/500 μg

Recombinant protein

Recombinant Human BMP-4 Protein


10 μg/100 μg/500 μg

Recombinant protein

Recombinant Human bFGF/FGF-2 Protein


10 μg/100 μg/500 μg

Auxiliary components for cell culture

Recombinant Human Insulin


25 mg/100 mg/500 mg/1 g


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