There is no known therapy for treatment of prion diseases, but recently a lot of new principles of treatment have been discovered. One of these are anti-prion antibodies which could prevent spreading of PrPSc. Mesenchymal stem cells (MSC) are potentially a delivery system for these antibodies to the brain, due to their self-renewal, differentiation, immunomodulation properties, pathotropism and ability to cross the blood-brain barrier. In this thesis, we attempted to insert a molecular construct for scFv fragment of anti-prion antibody V5B2 into mouse MSC by electroporation. In the first part, we optimized electroporation parameters, by using 8 square wave pulses, 1 ms long, with 1,0 kv/cm electric field strength and 1 Hz frequency. Cells were resuspended in high conductivity electroporation buffer to a final concentration of 3 x 106 cells/ml and 30 µg/ml of GFP plasmid was added. Electroporation was repeated with plasmids containing intracellular and secretory scFv V5B2. The cells were characterized by flow cytometry where, after electroporation, only a small difference in phenotype of mouse MSC was observed. We tried to determine the expression of scFv V5B2 in and outside of cells by immunocytochemistry and ELISA, but were not successful. According to our results, the scFv V5B2 is not expressed in mouse MSC. Further optimization of transfection methods, possible replacement of vectors and optimization of analytical methods for detection of expression is required.
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