Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
Zenzmaier, C.
Investigation of the proteome of CD34+ stem/progenitor cells from umbilical cord blood
[ Dissertation ] Medical University of Graz; 2004. pp.
- Autor*innen der Med Uni Graz:
- Betreuer*innen:
- Preisegger Karl Heinz
- Altmetrics:
- Abstract:
- Umbilical cord blood (UCB) has been used successfully as an alternative source of haematopoietic stem/progenitor cells (HSPC) in stem cell transplantation for the treatment of acquired and genetic diseases. Besides the many advantages of UCB derived stem cells there are two major disadvantages: First, the limited cell number, and second, the delayed haematopoietic recovery after transplantation. One concept to overcome these shortcomings is the ex-vivo expansion of cord blood stem cells to provide a stem cells pool, that allows multiple application of one single sample in one or several patients either for haematopoietic recovery or for tissue repair/regeneration. To perform latter, it can be beneficial to pre-differentiate the cells in-vitro cultures. In order to meet all required safety standards, the changes in the protein expression pattern of the cells during culture may serve as markers fo a desired or undesired differentiation pathway. The major aim of this thesis was to establish suitable proteomic methods for the investigation of the human stem cell proteome to create a master protein expression pattern for further comparison with expanded and/or differentiated cells. Fir this purpose, two dimensional polyacrylamide gel electrophoreses of five CD34+ preparations were performed and the protein patterns were compared for matching protein spots. Approximately only 17.5% of the proteins were found at identical positions in all samples. The identity of 22 of these protein spots was determined after excision and tryptic in-gel digestion of each protein spot by applying nano-LC coupled to ion trap MS/MS detection. In a second approach, the entire followed by two dimensional nano-LC and ion trap MS/MS detection. By this means, 215 proteins were reliably identified with a mowse score >80. In order to support the proteomic data additional RE-PCR and western blotting analyses were performed.