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Protein-Profiling analysis: Study of the influence of radiation on human embryonic stem cells

Eitel, T. (2017).

Master-Thesis, Course of Studies: Biomedical Sciences, Faculty of Life Sciences, Hochschule Albstadt-Sigmaringen. 93 Seiten.

Human embryonic stem cells are pluripotent cells which develop early during embryogenesis. Damages occurring from ionizing radiation like X- or Y-rays in these pluripotent stem cells have more profound effects than damages arising in adult cells, since defects in pluripotent stem cells are passed on to a large number of resulting differentiated cells. Neural cells from the central nervous system inherit damages which can lead to severe neurological or neurodegenerative diseases like Alzheimer's disease or tumors of the brain.

In this Master's thesis the effects of damaging radiation were addressed at two different time points during cell differentiation. First the human embryonic stem cell line H9 was differentiated to neuroepithelial progenitor cells (NEP) and further to neurospheres (NS). NEPs were treated with distinct doses (0.5 Gy and 1Gy) of Roentgen radiation. Protein expression was measured in these cells and compared with an unirradiated control cell culture. In a second experiment, H9 cells were irradiated as described and samples were collected one and two days after radiation. Changes in protein concentration induced by radiation were detected by employing a newly developed method, the DigiWest. This method allowed the semiquantitative detection of hundreds of proteins and generated a broad picture of changes in protein expression could be generated.

Significant variations of protein concentrations induced due to ionizing radiation was examined. For covering the biological range, proteins of several cellular pathways and morphological conditions were selected. Several proteins were detected showing changes in protein concentration with higher radiation doses. Most promising was p53, showing an increase with higher doses of radiation and a decrease in further differentiated cells.