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Types of Angiogenesis Assays

Tube Formation Assay

Cells seeded onto gel matrices can be used for monitoring their ability to form new vessels. A tube formation assay is performed by first seeding single cells, and then observing their characteristic patterns. Using the ibidi µ-Slide Angiogenesis or the μ-Plate Angiogenesis 96 Well allows you to investigate angiogenesis in tube formation assays.

Some possible experimental endpoints are:

  • the angiogenic potential of substances
  • the effect of inhibitors or enhancers on tube formation assays
  • molecular mechanisms visualized by fluorescence microscopy
  • investigation of signal transduction or cytoskeletal effects

More information:

In our Application Notes

you will find detailed information on how to setup and optimize tube formation experiments, data analysis, and interpretation.

Fluorescence microscopy of one single strand composed of HUVEC cells. F-actin cytoskeleton stained green, and cell nuclei blue.

Influence of inhibitors on tube formation over time. Please note the time dependence of the tube formation process and take images at identical time points.

Sprouting Assay

To carry out a sprouting assay, either spheroids or pieces of tissue, such as from the aorta, need to be placed on the gel matrix. The sprouting assay, like the tube formation assay, needs a well-defined thickness of the gel layer underneath, which is available with the µ-Slide Angiogenesis.

It is, therefore, a great benefit to perform these assays using the µ-Slide Angiogenesis. In addition to reproducible cell culture conditions, all cells are also placed in one optical plane.

3D Cell Cultures

The “well in a well“ feature of the µ-Slide Angiogenesis also supports the microscopy of cells embedded in gel matrices. 3D cell cultures mimic in vivo conditions, such as those of cancer cells and hepatocytes. Also, non-adherent cells, such as blood cells or bacteria, can be immobilized for enhanced microscopy access.

The amounts of gel matrix and the medium volume are balanced to create a nutrient supply for 3D cell cultures that have low and medium cell densities.

Application example:
A cell spheroid (or micro tumor) is cultivated on top of, or embedded in, a 3D matrix for long-term cultivation. The upper well provides nutrients by means of diffusion.