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1Department of Ophthalmology,University Hospital RWTH Aachen, 2Experimental Ophthalmology,University of Geneva, 3Department of Ophthalmology,University Hospitals of Geneva, 4Université de Paris, CNRS, INSERM, UTCBS, Unité des technologies Chimiques et Biologiques pour la Santé, 5Chimie ParisTech,PSL Research University, 6Max Delbrück Center for Molecular Medicine in the Helmholtz Association, 7Division of Medical Biotechnology,Paul-Ehrlich-Institute
Chapters
- 00:05Introduction
- 00:57Primary Human RPE Cell Electroporation and Plasmid DNA Preparation
- 01:57Cultivated Primary Human RPE Cell Preparation
- 03:19His-Tagged Pigment Epithelium-Derived Factor (PEDF) Fusion Protein Purification
- 04:57Results: Representative Primary Human RPE Transfection and Genetic Modification Evaluation
- 06:24Conclusion
Vi har utviklet en protokoll for å transfektere primære humane pigmentepitelceller ved elektroporering med genet som koder for pigmentepitelavledet faktor (PEDF) ved hjelp av Tornerose (SB) transposonsystemet. Vellykket transfeksjon ble demonstrert ved kvantitativ polymerasekjedereaksjon (qPCR), immunoblotting og enzymbundet immunosorbentanalyse (ELISA).
