{"id":{"repo_id":"freiburg-diss","oai_identifier":"oai:freidok.uni-freiburg.de:1683"},"canonical_url":"https://search.dev.ndltd.org/etd/freiburg-diss/oai:freidok.uni-freiburg.de:1683","repository":{"repo_id":"freiburg-diss","name":"University of Freiburg","base_url":"https://freidok.uni-freiburg.de/oai/oai2.php"},"display":{"title":"Gene transfer of keratinocytes with a human Factor XIII - plasmid construct","abstract":"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.","abstract_html":"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.","abstract_has_math":false,"creators":["Horváth, Laura"],"institution":null,"degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":["Stark, Gerhard Björn"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":null,"date_issued":"","date_published":null,"updated_at":"2026-07-24T02:22:22Z","subjects":["fibrin","blood clotting factor XIII","gene transfer","wound healing","transfection"],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://freidok.uni-freiburg.de/data/1683","outbound_label":"Repository record","outbound_source":"source_url"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Stark, Gerhard Björn"]},{"key":"dc:creator","label":"Author","values":["Horváth, Laura"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:type","label":"Dc Type","values":["DoctoralThesis"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["fibrin","blood clotting factor XIII","gene transfer","wound healing","transfection"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["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.","Faktor XIII (Plasmatransglutaminase) ist ein wichtiger Blutgerinnungsfaktor. Er katalysiert die Vernetzungen zwischen den Fibrinmonomeren, wodurch das Fibringerinnsel eine verbesserte Stabilität und einen erhöhten Widerstand gegen Fibrinolyse erreicht. Dieses liefert eine vorläufige Matrix für die Zellwanderung und Zelladhäsion und fördert dadurch die Wundheilung. Der Gentransfer bietet eine attraktive Methode für die lokalisierte und vorübergehend aktive Freisetzung von Faktoren zur Wundheilung an. Ziel dieser Studie war die transiente liposomale Transfektion von humanen Keratinozyten mit dem Blutgerinnungsfaktor XIII-Gen und die Analyse der Proteinproduktion für eine zukünftige in vivo Verwendung in der Wundheilung. Humane Keratinozyten wurden in vitro in serumfreiem Medium kultiviert. Bei 80% Konfluenz wurde mit unterschiedlichen Mischungen, bestehend aus DNA und EscortÒ Transfektionsreagenz, die liposomale Transfektion durchgeführt. Die Zellen wurden mit Hilfe des pCMX-FXIII Plasmids transfiziert. Der Überstand wurde 5 Tage lang täglich gesammelt. Die Proteinexpression wurde mit einem human FXIII spezifischen ELISA (Enzyme Linked Immunosorbent Assay) bestimmt. Humane Keratinozyten wurden erfolgreich mit Hilfe einer non-viralen Gentransfermethode transfiziert. Die Expression wurde über 5 Tage verfolgt. Die höchste Expression wurde an Tag 4 (158,26 ng/10.000 Zellen/ 24h) beobachtet. Nach unseren Kenntnissen dokumentieren diese Ergebnisse die erste erfolgreiche liposomale Transfektion von humanen Keratinozyten mit Faktor XIII. Diese Ergebnisse zeigen darauf hin, dass in vitro transfizierte Keratinozyten in der in vivo Produktion von Proteinen für die Therapie von nicht- heilenden Wunden eingesetzt werden könnten."]},{"key":"dc:format.medium","label":"Dc Format Medium","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Gene transfer of keratinocytes with a human Factor XIII - plasmid construct","Gentransfer von Keratinozyten mit einem humanen Faktor XIII - Plasmid Konstrukt"]}]}],"canonical_facts":{"dc:contributor":["Stark, Gerhard Björn"],"dc:creator":["Horváth, Laura"],"dc:description.abstract":["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.","Faktor XIII (Plasmatransglutaminase) ist ein wichtiger Blutgerinnungsfaktor. Er katalysiert die Vernetzungen zwischen den Fibrinmonomeren, wodurch das Fibringerinnsel eine verbesserte Stabilität und einen erhöhten Widerstand gegen Fibrinolyse erreicht. Dieses liefert eine vorläufige Matrix für die Zellwanderung und Zelladhäsion und fördert dadurch die Wundheilung. Der Gentransfer bietet eine attraktive Methode für die lokalisierte und vorübergehend aktive Freisetzung von Faktoren zur Wundheilung an. Ziel dieser Studie war die transiente liposomale Transfektion von humanen Keratinozyten mit dem Blutgerinnungsfaktor XIII-Gen und die Analyse der Proteinproduktion für eine zukünftige in vivo Verwendung in der Wundheilung. Humane Keratinozyten wurden in vitro in serumfreiem Medium kultiviert. Bei 80% Konfluenz wurde mit unterschiedlichen Mischungen, bestehend aus DNA und EscortÒ Transfektionsreagenz, die liposomale Transfektion durchgeführt. Die Zellen wurden mit Hilfe des pCMX-FXIII Plasmids transfiziert. Der Überstand wurde 5 Tage lang täglich gesammelt. Die Proteinexpression wurde mit einem human FXIII spezifischen ELISA (Enzyme Linked Immunosorbent Assay) bestimmt. Humane Keratinozyten wurden erfolgreich mit Hilfe einer non-viralen Gentransfermethode transfiziert. Die Expression wurde über 5 Tage verfolgt. Die höchste Expression wurde an Tag 4 (158,26 ng/10.000 Zellen/ 24h) beobachtet. Nach unseren Kenntnissen dokumentieren diese Ergebnisse die erste erfolgreiche liposomale Transfektion von humanen Keratinozyten mit Faktor XIII. Diese Ergebnisse zeigen darauf hin, dass in vitro transfizierte Keratinozyten in der in vivo Produktion von Proteinen für die Therapie von nicht- heilenden Wunden eingesetzt werden könnten."],"dc:format.medium":["application/pdf"],"dc:subject":["fibrin","blood clotting factor XIII","gene transfer","wound healing","transfection"],"dc:title":["Gene transfer of keratinocytes with a human Factor XIII - plasmid construct","Gentransfer von Keratinozyten mit einem humanen Faktor XIII - Plasmid Konstrukt"],"dc:type":["DoctoralThesis"]},"updated_at":"2026-07-24T02:22:22Z"}