The lungs together with the heart are the core of the human body. These organs maintain the homeostasis and provide substantial elements for proper functioning of the organism. Any kind of damage or dysfunction to these organs, results in serious disorders and general imbalance of the organism. Most critical are defects in the endothelial barrier, which consist of the endothelial cells (ECs). The vascular system is padded with a single layer of ECs. Endothelial cell structure and functional integrity are essential for the maintenance of the vessel wall and the circulation. Endothelial cell injury, activation or dysfunction is a feature of many pathologic states, just to mention few like inflammation or effects on vascular tone.Regenerative medicine, including cell replacement therapy, is a promising alternative for disease treatments. The application of in vitro-generated (and if required) genetically amended cells could help to recover the function of damaged parts of a specific organ. One of the approaches is to establish or restore normal function of affected endothelial cells, by therapies based on embryonic stem cells (ESCs). However, generation of those cells is ethically debatable. Development of induced pluripotent stem cells (iPS) from somatic cells has emerged as a solution. This enables the generation of patient- and disease-specific iPS cells, which may produce therapeutic cell populations without immune rejection and moral dispute. Regenerative therapies seem to carry a great promise to treat endothelial dysfunction in respiratory and cardiovascular diseases.To investigate the sites of integration of ES cell-derived endothelial (progenitor) cells, new cell lines from murine ES cells using lentiviral (LV) transduction were generated. In order to achieve the aim it had been decided to test different promoter-resistance-gene combinations. Therefore, a set of vectors containing murine promoters: VE-Cadherin (vascular endothelial) or VEGFR2 (vascular endothelial growth factor) in connection with GFP (green fluorescent protein) and antibiotic resistance genes hygromycin B, neomycin and puromycin, were produced and validated. It was feasible to generate and screen a cell line within a few weeks.The newly established cell lines are suitable for monitoring endothelial differentiation and selection by means of antibiotic resistance. These experiments demonstrated that for an effective antibiotic selection of the desired cell type, further investigations are required. It was possible to produce murine iPS cell lines, containing the above-mentioned transgenes. Carefully selected and purified murine ECs subsets will be used for in vivo studies in tumor angiogenesis models in subsequent studies.
Verknüpfung zu Publikationen oder weiteren Datensätzen
