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Updated: Jun 4, 2025
Generation of Human Blood Vessel Organoids from Pluripotent Stem Cells
Published on: January 20, 2023
Protocol for efficient generation of human artery and vein endothelial cells from pluripotent stem cells
Kyle M Loh1, Sherry Li Zheng1, Kevin J Liu2
1Institute for Stem Cell Biology & Regenerative Medicine, Stanford University, Stanford, CA 94305, USA; Department of Developmental Biology, Stanford University, Stanford, CA 94305, USA.
Insights
This study details a rapid 3-4 day protocol for generating human artery and vein endothelial cells (ECs) from pluripotent stem cells. The method uses defined, serum-free media without genetic modification or feeder cells.
Area of Science:
- Stem cell biology
- Vascular biology
- Cell differentiation
Background:
- Blood vessels are crucial for organ function in both health and disease.
- Efficient generation of specific endothelial cell types is vital for research and therapeutic applications.
- Existing methods for endothelial cell generation can be time-consuming or require complex procedures.
Purpose of the Study:
- To present a protocol for the rapid and efficient differentiation of human pluripotent stem cells into artery and vein endothelial cells (ECs).
- To establish a method for generating ECs within a short timeframe (3-4 days).
- To provide a protocol that avoids feeder cells and genetic manipulation, simplifying the process.
Main Methods:
- Human pluripotent stem cells are cultured as monolayers in defined, serum-free media.
- Sequential differentiation involves inducing primitive streak and lateral mesoderm stages.
- Specific conditions are applied to direct differentiation towards either artery or vein endothelial cells.
Main Results:
- Successful generation of human artery and vein endothelial cells (ECs) from pluripotent stem cells.
- Differentiation achieved within a rapid 3-4 day period.
- Protocol demonstrated efficacy using defined, serum-free culture conditions without feeder cells or genetic manipulation.
Conclusions:
- The presented protocol offers an efficient and rapid method for generating human artery and vein endothelial cells (ECs).
- This streamlined approach is suitable for research applications requiring specific EC subtypes.
- The method's simplicity, using defined media and avoiding complex manipulations, enhances its accessibility.
Abstract:
Blood vessels permeate all organs and execute myriad roles in health and disease. Here, we present a protocol to efficiently generate human artery and vein endothelial cells (ECs) from pluripotent stem cells within 3-4 days of differentiation. We delineate how to seed human pluripotent stem cells and sequentially differentiate them into primitive streak, lateral mesoderm, and either artery or vein ECs. We differentiate stem cells in defined, serum-free culture media in monolayers, without feeder cells or genetic manipulations. For complete details on the use and execution of this protocol, please refer to Ang et al. 1.
