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Angiogenesis Invasion Assay to Study Endothelial Cell Invasion and Sprouting Behavior.

Yuechao Dong1, Florian Alonso1, Tiya Jahjah1

  • 1University Bordeaux, INSERM, Centre de Recherche cardio-thoracique de Bordeaux, U1045, Bordeaux, France.

Methods in Molecular Biology (Clifton, N.J.)
|January 18, 2023
PubMed
Summary

This study introduces a new in vitro assay to model angiogenesis, the formation of new blood vessels. The assay effectively simulates the invasion step, enabling research into blood vessel formation and drug screening.

Keywords:
AngiogenesisBasement membraneCell invasionEndothelial cellsFluorescence microscopyIn vitro angiogenesis assayNotch signalingPodosomesTip cellsVascular sprout

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Area of Science:

  • Biomedical Engineering
  • Cell Biology
  • Developmental Biology

Background:

  • Angiogenesis, the formation of new blood vessels, is crucial for development but also implicated in various diseases.
  • This complex process involves endothelial cell activation, migration, and tube formation, culminating in vessel anastomosis.
  • Existing models often fail to fully replicate the in vivo invasion step of angiogenesis.

Purpose of the Study:

  • To develop and describe a novel in vitro assay that accurately mimics the invasion step of angiogenesis.
  • To provide a tool for studying physiological and pathological angiogenesis.
  • To enable high-throughput screening of compounds for pro-angiogenic or anti-angiogenic properties.

Main Methods:

  • Human microvascular endothelial cells were cultured in a three-dimensional, VEGF-enriched basement membrane environment.
  • The assay was designed to promote endothelial cell sprouting, tube formation, and anastomosis.
  • Vascular sprouts and endothelial cell behavior were observed using various microscopy techniques.

Main Results:

  • The assay successfully induced endothelial cell sprouting and tube formation within hours.
  • Endothelial cells adopted tip cell characteristics, expressing relevant markers and exhibiting dynamic behavior.
  • The in vitro model demonstrated cell dynamics comparable to in vivo angiogenic processes.

Conclusions:

  • The developed assay provides a robust and manipulable system for studying the invasion phase of angiogenesis.
  • This model facilitates genetic manipulation for in-depth research into angiogenesis mechanisms.
  • The assay serves as a valuable platform for screening potential therapeutic compounds targeting angiogenesis.