Knockdown von Runx2 durch RNA-Interference in chondrogenen Progenitor-Zellen
Knockdown of Runx2 with RNA-interference in chondrogenic progenitor cells
von Moritz Klein
Datum der mündl. Prüfung:2014-03-26
Erschienen:2014-03-26
Betreuer:Prof. Dr. Nicolai Miosge
Gutachter:Prof. Dr. Nicolai Miosge
Gutachter:Prof. Dr. Dr. Karl Günter Wiese
Dateien
Name:ENDE Gedruckt 20.03.14 1 Diss Top Kopie.pdf
Size:20.0Mb
Format:PDF
Zusammenfassung
Englisch
The regeneration of diseased hyaline cartilage continues to be a great challenge because its self renewal capacity is limited. In repair tissue from human articular cartilage during the late stages of osteoarthritis chondrogenic progenitor cells (CPC`s) were found. These cells showed stemm cell capacity such as multipotency and migratory activity. The Transkriptionfaktor Runt-related transkription Factor 2 (Runx2) is an importend regulator of Osteogenesis and is also expressed in CPC`s. We tried to generate a Knockdown of Runx2 in CPC`S with the help of an lentiviral Vektor and RNA-Interference. We also investigated the influence of Runx2 knockdown to the chondrogenic transkriptionfactor SRY-related HMG-box gene 9 (Sox9). Our results showed that the transfection of CPC`s with the i-lenti-GFP expression Vector (Biocat) with Amaxa Nukleofection leads to transient transfektion. The Knockdown of Runx2 could be showed to 96 h after transfektion with Polymerase chain reaction (RT-PCR) and to 48 h in the western blot. We also could show a changing in sox9 expression in the transfected CPC`s.
Keywords: Runx2; Sox9; Osteoarthritis; RNA-interference; chondrogenic progenitorcells; Nucleofaction