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Updated: Mar 7, 2026

Generation of Human Blood Vessel Organoids from Pluripotent Stem Cells
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Animal-origin-free method for generating blood vessel organoids.

Alexander Hoffmann1,2, David Schorn1, Jakob Thönig1

  • 1Angios FlexCo, Exlgasse 24, 6020, Innsbruck, Austria.

Scientific Reports
|March 5, 2026
PubMed
Summary

We developed a simplified, animal-origin-free method for generating blood vessel organoids (BVOs) using a novel "sitting drop" culture technique. This approach enhances reproducibility and automation for vascular disease modeling and regenerative therapies.

Keywords:
BioengineeringBlood vessel organoids (BVO)CollagenExtracellular matrix (ECM)High-throughputOrganoids

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Area of Science:

  • Biotechnology
  • Regenerative Medicine
  • Vascular Biology

Background:

  • Blood vessel organoids (BVOs) are crucial for studying vascular diseases and developing therapies.
  • Current BVO generation methods are complex, labor-intensive, and use animal-derived materials, hindering reproducibility and clinical use.

Purpose of the Study:

  • To develop a simplified, animal-origin-free protocol for BVO generation.
  • To enable high-throughput automated workflows for BVO production.
  • To improve reproducibility and scalability of BVO technology for clinical translation.

Main Methods:

  • Utilized ultra-low attachment 96-well U-bottom plates for standardized human induced pluripotent stem cell (hiPSC) aggregation.
  • Developed a single-layer
  • sitting drop
  • culture method using a human-derived collagen-based extracellular matrix.
  • Employed flow cytometry to analyze cellular composition and in vivo transplantation to assess functional integration.

Main Results:

  • Generated BVOs with reproducible morphology and cellular composition, including CD31+ endothelial cells and PDGFRβ+ pericytes.
  • The
  • sitting drop
  • method requires fewer materials and handling steps compared to traditional two-layer cultures.
  • Transplanted BVOs integrated into host vasculature in a mouse wound model, demonstrating therapeutic potential.

Conclusions:

  • The novel
  • sitting drop
  • protocol offers a robust, animal-origin-free, and automation-ready method for BVO generation.
  • This simplified approach enhances reproducibility and scalability, facilitating high-throughput screening and clinical applications.
  • The developed method paves the way for advancing vascular organoid technology in research and therapeutics.