A. Shakhbazau, D. Shcharbin, M. Bryszewska, R. Kumar, H. M. Wobma, M. S. Kallos, N. Goncharova, I. Seviaryn, S. Kosmacheva, M. Potapnev and R. Midha Pages 5572 - 5579 ( 8 )
Genetic engineering of stem cells and their derivatives has the potential to enhance their regenerative capabilities. Here, dendrimer- and lipofection-based approaches were used for non-viral neurotrophin-3 (NT-3) over-expression in Schwann cells differentiated from skin precursors (SKP-SCs). A variety of dendrimers were first tested for transfection efficiency on HEK 293T cells, with PAMAMNH2 G4 found most effective and used subsequently for SKP-SCs transfection. Plasmid-based expression resulted in increased NT-3 release from SKP-SCs in both adherent and microcarrier-based culture. In a proof-of-concept study, the microcarrier/SKP-SCs were implanted into the injured nerve, and transfected cells were shown to detach, integrate into the nerve tissue and associate with regenerating axons. Virus-free systems for transient neurotrophin expression are a feasible and biologically safe option to increase the therapeutic value of stem cells and stem cell-derived cells in nerve repair strategies. Further work to develop bioprocesses for expansion of SKP-SCs on microcarriers in bioreactors is still needed.
Skin precursor-derived Schwann cells, NT-3, transfection, non-viral gene delivery, dendrimers, PAMAM, lipofectamine, microcarriers, cytodex 3, peripheral nerve
Department of Clinical Neuroscience, University of Calgary, HMRB 109-3330 Hospital Drive NW T2N4N1, Calgary, Canada.