Directed In Vivo Angiogenesis Assay

The Directed In Vivo Angiogenesis Assay (DIVAA™) is the first in vivo system for the study of angiogenesis that provides quantitative and reproducible results. Angioreactors (silicone cylinders closed at one end) containing 20 µL of basement membrane with/without angiogenic-modulating factors are implanted subcutaneously in the dorsal flank of nude mice. With the onset of angiogenesis, vascular endothelial cells proceed to grow into the basement membrane extract and form vessels in the angioreactor. As early as nine days post-implantation, there are enough cells to determine an effective dose response to angiogenic modulating factors.

DIVAA™ has been used with nude mice, the recommended and qualified strain for the assay. The mouse strain C57Bl has reportedly been used successfully with the kit. However, optimization is required, and variability is commonly reported. If you require C57BI mice for your experiments, it is recommended to allow the implantation to go for up to 15 days.

Yes, the materials supplied are tested following USP <71> sterility guidelines.

While FGF-2 and VEGF have both been demonstrated to promote angiogenesis in DIVAA™, the FGF-2/VEGF mixture provides a synergistic effect allowing a drastic increase in response using less total growth factor.

FITC-Lectin specifically binds to endothelial cells, so it counts the total number of endothelial cells contained within the angioreactor. FITC-Dextran does not bind endothelial cells. Instead, it is dispersed within the blood of the mouse, being evenly distributed within the blood vessels, so it quantitates the total volume of blood contained within the vessels within the angioreactor. Both procedures have demonstrated equivalent results empirically.

The angioreactors should be implanted on the host mouse dorsal lateral flanks. These areas exhibit sufficient vasculature to support initiation of angiogenesis, and they are also strategically positioned to prevent removal by the host mouse. Other areas may be used for implantation; however, this is outside of the product specification. Please consult the scientific literature for more information.

While some groups have had success with implanting angioreactors containing viable cells, this application has not been qualified by R&D Systems. Please consult the scientific literature for more information.

The DIVAA kit has been evaluated using nude mice.  Although C57Bl/6 mice can be used for this assay, we recommend the use of nude mice for optimal results.

Growth of connective tissue into the angioreactor during angiogenesis is normal. The amount of connective tissue is dependent on the strain of mice, treatment, and response. The cell dissociation step may be increased up to 3 hours to improve dissociation/degradation, and any remaining connective tissue may be removed. If this does not solve the problem, it may be more appropriate to use the Optional Protocol for Dextran-FITC Detection (which measures capillary volume and discards cellular/connective tissue debris).