Non-viral engineering of skin precursor-derived Schwann cells for enhanced NT-3 production in adherent and microcarrier culture

Curr Med Chem. 2012;19(32):5572-9. doi: 10.2174/092986712803833218.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cells, Cultured
  • Dendrimers
  • Female
  • HEK293 Cells
  • Humans
  • Nerve Regeneration
  • Neurotrophin 3 / metabolism*
  • Polypropylenes
  • Rats
  • Rats, Inbred Lew
  • Schwann Cells / metabolism*
  • Sciatic Nerve / injuries
  • Sciatic Nerve / physiology
  • Skin / cytology
  • Stem Cell Transplantation
  • Transfection / methods*

Substances

  • Dendrimers
  • Neurotrophin 3
  • PAMAM Starburst
  • Polypropylenes
  • poly(propyleneimine)