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Preparation for external prevalidation: Optimization of the internally validated and standardized method for quantitation of angiogenesis and antiangiogenesis in vitro by determination of the angiogenic standard of the cells to be used
Project
Project code: BfR-ZEBET-08-1328-206
Contract period: 01.08.2007
- 31.07.2010
Purpose of research: Applied research
The aim of the project is the optimization of an internally validated and standardized method for quantitation of angiogenesis and antiangiogenesis in vitro. This method is based on in vitro cultivation of microvascular endothelial cells. The cells were stimulated by growth media and showed characteristic morphological changes, which could be assigned to the phases of the angiogenic cascade in vivo and were divided into strictly defined stages. Quantitation of angiogenesis and antiangiogenesis was carried out via time-dependent ascertainment of the defined stages. This project focuses on the endothelial cells used within the established method. Therefore, the protein expression patterns of four human endothelial cell cultures, which only in two cases could be stimulated to run chronologically through all stages of angiogenesis, should be analyzed by two-dimensional gel electrophoresis. These proteins exclusively and similarly expressed in the angiogenic cultures shall be determined by comparing the protein expression patterns. Subsequently, suppression of the expression of these proteins by gene silencing will show which proteins are responsible for the angiogenic potential of endothelial cells. In future these proteins can be used as markers to prove the angiogenic potential of endothelial cell cultures and therefore to verify their applicability for quantitation of angiogenesis and antiangiogenesis in vitro. These investigations will provide the pre-requisites for the prevalidation of the established method for quantitation of angiogenesis and antiangiogenesis in vitro in external labs.
Section overview
Subjects
- Animal health
- Biotechnology
Framework programme
Funding programme
Excutive institution
BfR - Centre for Documentation and Evaluation of Alternatives to Animal Experiments (BfR - ZEBET)