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: Skin precursor-derived Schwann cells, NT-3, transfection, non-viral gene delivery, dendrimers, PAMAM, lipofectamine, microcarriers, cytodex 3, peripheral nerve
Current Medicinal Chemistry
Title:Non-Viral Engineering of Skin Precursor-Derived Schwann Cells for Enhanced NT-3 Production in Adherent and Microcarrier Culture
Volume: 19 Issue: 32
Author(s): 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
Affiliation:
Keywords: Skin precursor-derived Schwann cells, NT-3, transfection, non-viral gene delivery, dendrimers, PAMAM, lipofectamine, microcarriers, cytodex 3, peripheral nerve
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.
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Shakhbazau A., Shcharbin D., Bryszewska M., Kumar R., M. Wobma H., S. Kallos M., Goncharova N., Seviaryn I., Kosmacheva S., Potapnev M. and Midha R., Non-Viral Engineering of Skin Precursor-Derived Schwann Cells for Enhanced NT-3 Production in Adherent and Microcarrier Culture, Current Medicinal Chemistry 2012; 19 (32) . https://dx.doi.org/10.2174/092986712803833218
DOI https://dx.doi.org/10.2174/092986712803833218 |
Print ISSN 0929-8673 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-533X |
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