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Non-Viral Engineering of Skin Precursor-Derived Schwann Cells for Enhanced NT-3 Production in Adherent and Microcarrier Culture

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Abstract:

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.

Keywords: NT-3; PAMAM; Skin precursor-derived Schwann cells; cytodex 3; dendrimers; lipofectamine; microcarriers; non-viral gene delivery; peripheral nerve; transfection

Document Type: Research Article

DOI: https://doi.org/10.2174/092986712803833218

Publication date: 2012-11-01

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