Determining of the factors necessary for the growth, proliferation and survival of vascular endothelial cells and the functions of these factors

Angiogenesis is the process of forming new vessels through budding of capillary and vascular networks from existing vessels. The development of the formed vascular networks is determined by the presence of the basic factors necessary for the healthy growth of the cells around the vascular structure. Angiogenesis is a very complex mechanism due to the involvement of different factors in the process and the need to provide appropriate concentrations of these factors. The basic components of angiogenesis are defined as growth factors, cytokines and cytokine receptors that trigger different properties. Pericytes, which form the capillary wall together with endothelial cells, are of great importance. The mixture containing the factors used in the angiogenesis process should be in the right ratio and the best candidate that can produce this is macrophage cells. In the study, mammalian macrophage cells (J774.2) stored at -80 ̊C were dissolved with the help of a hot water bath. A medium containing Roswell Park Memorial Institute Media (RPMI)+ 10% FBS 1%+ 1% antibiotic [100 μg/MI pecillin and 100 μg/Ml streptomycin] was added to mammalian macrophage cells (J774-2) at different rates, and 100 cm2 It was kept in the cell culture plate until it reached 90% confluency (covering 90% of the culture dish). Protein types and amounts were investigated by proteomic analysis in order to determine the factors secreted by mammalian macrophage cells and their amounts in vitro. According to the results of proteomic analysis, the taxonomy of macrophage cells was confirmed as (J774-2) mus musculus. In addition, medium addition (20 μl, 40 μl, 60 μl, 80 μl and 100 μl) was made at 24-hour, 48-hour and 72-hour intervals during MTT analysis on HUVEC cells. In the study, % cell growth rates were evaluated at absorbance at 570 nm using detergent reagent (SDS). Follow-up of the effects of factors that increase the formation of vessels was carried out by taking images with a fluorescent microscope within the scope of the study. The effect of HUVEC cells on vessel formation is carried out in two stages as lyophilized and normal conditions, and its contribution to extending the shelf life from 6 months to 18 months is investigated.