Gene transfer of keratinocytes with a human Factor XIII - plasmid construct

Factor XIII (plasma transgutaminase) is essential to the clotting process. It catalyses the cross-links between the fibrin monomers, in consequence of the forming fibrin clot achieves an improved stability and a high fibrinolysis resistance. This provides a provisional matrix for the cell migration and attachment, thereby contributes to wound healing. Gene transfer offers an attractive method for localised and transient active delivery of factors to healing wounds. Aim of this study was the transient liposomal transfection of human keratinocytes with the gene for clotting factor XIII and the analysis of the protein expression for a future in vivo use for wound healing. Human keratinocytes were cultured in vitro with serum free medium. At 80% confluence liposomal transfection was performed using different DNA-EscortÒ transfection reagent mixtures. Cells were transfected using a pCMX-FXIII plasmid. The supernatant was collected each day for 5 days. Protein expression was determined with a specific ELISA (Enzyme Linked Immunosorbent Assay) for human FXIII. Human keratinocytes were successfully transfected using a non-viral gene transfer method, the expression was detected over 5 days. Highest expression was detectable on day 4 (158,26 ng/10.000 cells/ 24h). To our knowledge the results document the first successful liposomal transfection of human keratinocytes with FXIII. These data suggest the possibility to use in vitro transfected keratinocytes for the in vivo production of therapeutic proteins for the therapy of non-healing wounds.

Author dc:creator

• Horváth, Laura

Contributors dc:contributor

• Stark, Gerhard Björn

• fibrin
• blood clotting factor XIII
• gene transfer
• wound healing
• transfection

Repository record source_url

https://freidok.uni-freiburg.de/data/1683

OAI identifier oai:identifier

oai:freidok.uni-freiburg.de:1683