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Updated: Jun 17, 2026
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In Vitro Three-Dimensional Sprouting Assay of Angiogenesis Using Mouse Embryonic Stem Cells for Vascular Disease Modeling and Drug Testing
Published on: May 11, 2021
Engineering surfaces for site-specific vascular differentiation of mouse embryonic stem cells
C Katherine Chiang1, Mohammad Fahad Chowdhury, Rohin K Iyer
1Institute of Biomaterials and Biomedical Engineering, University of Toronto, Ont., Canada.
Acta Biomaterialia
|December 17, 2009
Summary
Immobilizing vascular endothelial growth factor-A (VEGF) on surfaces guides stem cell differentiation. This method spatially controls stem cell fate, promoting either endothelial or smooth muscle cell development based on VEGF presence.
Area of Science:
- Biotechnology
- Stem Cell Biology
- Tissue Engineering
Background:
- Stem and progenitor cell differentiation typically uses soluble growth factors, offering temporal but not spatial control.
- Angiogenic progenitor cells are crucial for blood vessel formation.
Purpose of the Study:
- To investigate the spatial control of stem cell differentiation using patterned immobilized vascular endothelial growth factor-A (VEGF).
- To differentiate angiogenic progenitor cells derived from mouse embryonic stem cells (ESCs) on defined surface patterns.
Main Methods:
- Mouse ESCs expressing eGFP under Flk1 promoter were used to isolate Flk1+ angiogenic progenitors via fluorescence-activated cell sorting.
- Vascular endothelial growth factor-A (VEGF) was immobilized onto collagen IV (ColIV) using EDC chemistry.
- VEGF-ColIV lanes were stamped onto a photocrosslinkable chitosan layer, and Flk1+ progenitors were seeded for site-specific differentiation.
Main Results:
- Cultivation on immobilized VEGF surfaces yielded primarily endothelial cells (53% CD31 positive).
- Surfaces without immobilized VEGF favored vascular smooth muscle-like cell differentiation (38% smooth muscle actin positive).
- Controlled differentiation into distinct cell lineages was achieved through surface patterning.
Conclusions:
- Immobilized VEGF gradients provide spatial cues for directed stem cell differentiation.
- This technique offers a novel approach for tissue engineering and regenerative medicine applications.
- Surface-bound growth factors enable precise control over cell fate decisions.
