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Angiogenesis

, Volume 20, Issue 4, pp 493–504 | Cite as

Low levels of physiological interstitial flow eliminate morphogen gradients and guide angiogenesis

  • Venktesh S. Shirure
  • Andrew Lezia
  • Arnold Tao
  • Luis F. Alonzo
  • Steven C. GeorgeEmail author
Original Paper

Abstract

Convective transport can significantly distort spatial concentration gradients. Interstitial flow is ubiquitous throughout living tissue, but our understanding of how interstitial flow affects concentration gradients in biological processes is limited. Interstitial flow is of particular interest for angiogenesis because pathological and physiological angiogenesis is associated with altered interstitial flow, and both interstitial flow and morphogen gradients (e.g., vascular endothelial growth factor, VEGF) can potentially stimulate and guide new blood vessel growth. We designed an in vitro microfluidic platform to simulate 3D angiogenesis in a tissue microenvironment that precisely controls interstitial flow and spatial morphogen gradients. The microvascular tissue was developed from endothelial colony forming cell-derived endothelial cells extracted from cord blood and stromal fibroblasts in a fibrin extracellular matrix. Pressure in the microfluidic lines was manipulated to control the interstitial flow. A mathematical model of mass and momentum transport, and experimental studies with fluorescently labeled dextran were performed to validate the platform. Our data demonstrate that at physiological interstitial flow (0.1–10 μm/s), morphogen gradients were eliminated within hours, and angiogenesis demonstrated a striking bias in the opposite direction of interstitial flow. The interstitial flow-directed angiogenesis was dependent on the presence of VEGF, and the effect was mediated by αvβ3 integrin. We conclude that under physiological conditions, growth factors such as VEGF and fluid forces work together to initiate and spatially guide angiogenesis.

Keywords

αvβ3 integrin Concentration gradients Microphysiological systems Organ-on-a-chip 

Notes

Acknowledgements

This work was supported by grants from the National Institutes of Health (UH3 TR00048, R01 CA170879, and F31 CA163049). We would like to thank Mr Vinson Tran, Mo Kebaili, Linda McCarthy (University of California, Irvine) and Sandra Lam (Washington University in St. Louis) for technical assistance. We would also like to thank Dr. Abraham Lee for helpful advice and material assistance during the development stages of this project.

Supplementary material

10456_2017_9559_MOESM1_ESM.docx (16 kb)
Supplementary material 1 (DOCX 15 kb)
10456_2017_9559_MOESM2_ESM.pptx (11.4 mb)
Supplementary material 2 (PPTX 11692 kb)

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Copyright information

© Springer Science+Business Media Dordrecht 2017

Authors and Affiliations

  • Venktesh S. Shirure
    • 1
  • Andrew Lezia
    • 1
  • Arnold Tao
    • 1
  • Luis F. Alonzo
    • 2
  • Steven C. George
    • 1
    • 3
    Email author
  1. 1.Department of Biomedical EngineeringWashington University in St. LouisSt. LouisUSA
  2. 2.Department of Biomedical EngineeringUniversity of CaliforniaIrvineUSA
  3. 3.Department of Energy, Environment, and Chemical EngineeringWashington University in St. LouisSt. LouisUSA

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