When individual tumor cells grow into a macroscopic tumor they drastically increase their demand for oxygen and nutrients. An important example of angiogenesis is therefore the generation of new blood vessels to feed a tumor. In vitro angiogenesis assays will provide better understanding of these processes on a cellular and molecular level.
The µ-Slide Angiogenesis was designed for biomedical and pharmaceutical research. It can be used with all common gel matrices, like Matrigel®, collagen gels, and hyaluronic acid gels. Only 10µl gel per well are needed.
Cells on gel matrices can be used to monitor their ability to form new vessels. The tube formation assay is done by seeding single cells and observing charac-teristic patterns. In case of the sprouting assays either spheroids or pieces of tissue e.g. from aorta are placed on the gel matrix. All these assays share a need for a well defined thickness of the gel layer underneath.
The assays performed on the u-Slide Angiogenesis benefit from a well defined thickness of the gel matrices. Besides reproducible cell culture conditions, the cells are placed in one optical plane.
The u-Slide Angiogenesis was primarily developed to simplify the tube formation assay of endothelial cells on Matrigel®. This kind of assay is usually done in multiwell plates. In large wells as in the 6- or 12-well plates the required amount of gel matrix is large and very expensive. A barrier for going to smaller wells is the strong meniscus formation of the gel surface, which leads to inhomogenous cell distribution patterns. And even worse, cells on the bent gel surface are never in one optical plane. Another meniscus is formed at the air-liquid interface which restricts phase contrast only to a very small area at the center of the well.
The u-Slide Angiogenesis solves these problems providing a 4 mm well in a 5 mm well. This optical improvement is called “well in a well“.
Filling the smaller well with exactly 10µl of gel results in a plane gel surface. Filling the major well with exactly 50µl avoids meniscus formation also on the air-liquid interface.
The “well in a well“ feature of the u-Slide Angiogenesis also supports the microscopy of cells embedded in gel matrices. 3D cell cultures mimick in vivo conditions e.g. of cancer cells and hepatocytes. Also non adherent cells like blood cells or bacteria can be immobilzed for enhanced microscopy access.
The used amounts of gel matrix and medium volume are balanced for the nutrient supply of those 3D cell cultures with low and medium cell densities.