Protocols

Introduction

The cornea is an ideal model for studying in vivo angiogenesis since it is avascular and therefore any vascular development is the result of introduced material. Originally developed for rabbit eyes, it has also been adapted for mice, which are more widely used.¹,²

Assay Description

The assay is carried out by first creating a pocket in the cornea where the compound of interest is inserted. A variety of material can be used to hold the test cells containing angiogenic compounds in suspension, including sponges, ethylene vinyl copolymer, or Hydron. The vascular response can then be monitored by direct observation using either a stereomicroscope or slit lamp.

It is also possible to quantify the amount of vascularization by measuring the area of vessel penetration or by employing fluorescence techniques. Advantages of this assay include the ability to monitor progress of vessel formation, the absence of existing background vasculature, and adaptation for use in mice.  Disadvantages to consider include the need to perform a demanding surgical procedure which imposes limitations on the size of the study and the development of inflammation is common and may interfere with visualization.

Materials

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References

  1. Hartwell D, Butterfield C, Frenette P et al. (1998). Angiogenesis in P- and E-selectin-deficient mice. Microcirculation. 5(2-3):173-178.
  2. Presta M, Rusnati M, Belleri M et al. (1999). Purine analogue 6-methylmercaptopurine riboside inhibits early and late phases of the angiogenesis process. Cancer Res. 59(10):2417-2424.

LT172      updated  7-Oct-2011
 

For Research Use Only. Not for use in diagnostic procedures.