Evaluation of the angiogenesis process by endothelial cells

Introduction

Angiogenesis, the process by which endothelial cells generate new blood vessels derived from an existing vasculature, can be assessed by the ability of endothelial cells to form three-dimensional structures.

The formation of tubules by endothelial cells in an extracellular matrix mimicking the in vivo environment may be a relevant test to easily identify angiogenesis inhibitors.

Protocol

Format96 wells
Cellular model Human Microvascular Endothelial Cells-1 (ATCC CRL-3243) 
CoatingMatrigel
Reference molecule Suramin
Incubation24H at 37°C under 5% CO2 
Analysis

Incucyte® S3 analysis (SARTORIUS) - Cell imaging software
or ImageXpress Pico analysis (Molecular Devices)
→ Determination of the number of tubes
→ Determination of the length of tubes

Notes :

  • Allow 50 µL at 10 mM or 3 mg of sample to test 5 concentrations (3 points/concentration)
  • Cell viability is assessed in parallel

Recommendation :

  • Ensure solubility of the sample in a solvent compatible with cell culture

Inhibition of angiogenesis by Suramin

0 µM Suramin - 24h treatment
20 µM Suramin - 24h treatment
25 µM Suramin - 24h treatment
27.5 µM Suramin - 24h treatment
30 µM Suramin - 24h treatment
35 µM Suramin - 24h treatment