In a viscous medium endothelial cells (ECs) aggregate overnight to form a cellular spheroid. This behaviour was used to establish a spheroid-based cellular angiogenesis assay. For this assay human EC spheroids are embedded in a defined matrix (e.g. collagen or fibrin), are stimulated and finally EC sprouting is quantified.

Pro-angiogenic compounds/growth factors induce EC sprouting; anti-angiogenic compounds inhibit growth factor (e.g. VEGF-A, FGF-2) induced EC sprouting.

Steps of the cellular angiogenesis assay – EC spheroids are prepared by pipetting 500 ECs in a hanging drop on plastic dishes to allow spheroid aggregation overnight (left). EC spheroids are harvested and mixed with e.g. collagen which is allowed to polymerize in a 24-well plate (right). Test compounds are added and the plates are incubated for 24h. Assays are quantitatively analyzed by determining the cumulative sprout length per spheroid. For each compound concentration 10 spheroids are analyzed.

• The assay system is suitable for angiogenic/anti-angiogenic lead identification and optimization
• ProQinase offers single concentration testing of anti-angiogenic compounds and standard IC₅₀ determination of such compounds by testing at 7 concentrations in semi-log steps
• Assays for potential angiogenic compounds can be designed according to customers' need
  

The attractiveness of this test system for large scale screening is described in a poster, published 2007 at the AACR meeting.

Fact Sheet - Spheroid-Based Cellular Angiogenesis Assay

Cellular Phosphorylation Assays

Differential Cytotoxicity Assay

Spheroid-Based in vivo Angiogenesis Assay

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