title: Non-viral engineering of skin precursor-derived Schwann cells for enhanced NT-3 production in adherent and microcarrier culture.
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.
authors: Department of Clinical Neuroscience, Faculty of Medicine, University of Calgary, Calgary, Canada. shakhbazau@gmail.com
authors: A
authors: A
authors: Shakhbazau
authors: D
authors: D
authors: Shcharbin
authors: M
authors: M
authors: Bryszewska
authors: R
authors: R
authors: Kumar
authors: H M
authors: HM
authors: Wobma
authors: M S
authors: MS
authors: Kallos
authors: N
authors: N
authors: Goncharova
authors: I
authors: I
authors: Seviaryn
authors: S
authors: S
authors: Kosmacheva
authors: M
authors: M
authors: Potapnev
authors: R
authors: R
authors: Midha
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D050091:Dendrimers; D005260:Female; D057809:HEK293 Cells; D006801:Humans; D009416:Nerve Regeneration; D020933:Neurotrophin 3 / Q000378:metabolism*; D011126:Polypropylenes; D051381:Rats; D011917:Rats, Inbred Lew; D012583:Schwann Cells / Q000378:metabolism*; D012584:Sciatic Nerve / Q000293:injuries / Q000502:physiology; D012867:Skin / Q000166:cytology; D033581:Stem Cell Transplantation; D014162:Transfection / Q000379:methods*
chemical_list: D050091:Dendrimers; D020933:Neurotrophin 3; C104700:PAMAM Starburst; D011126:Polypropylenes; C443641:poly(propyleneimine)