Efficient transduction of equine adipose-derived mesenchymal stem cells by VSV-G pseudotyped lentiviral vectors

Res Vet Sci. 2014 Dec;97(3):616-22. doi: 10.1016/j.rvsc.2014.09.004. Epub 2014 Oct 6.

Abstract

Equine adipose-derived mesenchymal stem cells (EADMSC) provide a unique cell-based approach for treatment of a variety of equine musculoskeletal injuries, via regeneration of diseased or damaged tissue, or the secretion of immunomodulatory molecules. These capabilities can be further enhanced by genetic modification using lentiviral vectors, which provide a safe and efficient method of gene delivery. We investigated the suitability of lentiviral vector technology for gene delivery into EADMSC, using GFP expressing lentiviral vectors pseudotyped with the G glycoprotein from the vesicular stomatitis virus (V-GFP) or, for the first time, the baculovirus gp64 envelope protein (G-GFP). In this study, we produced similarly high titre V-GFP and G-GFP lentiviral vectors. Flow cytometric analysis showed efficient transduction using V-GFP; however G-GFP exhibited a poor ability to transduce EADMSC. Transduction resulted in sustained GFP expression over four passages, with minimal effects on cell viability and doubling time, and an unaltered chondrogenic differentiation potential.

Keywords: Adipose; Equine; Gene delivery; Green fluorescent protein; Lentiviral vector; Mesenchymal stem cells.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adipose Tissue / cytology*
  • Analysis of Variance
  • Animals
  • Baculoviridae / genetics
  • Cell Differentiation / physiology
  • DNA Primers / genetics
  • Genetic Therapy / methods*
  • Genetic Vectors / genetics*
  • Horse Diseases / genetics
  • Horse Diseases / therapy*
  • Horses
  • Lentivirus / genetics*
  • Mesenchymal Stem Cells / physiology*
  • Molecular Imaging / veterinary
  • Transduction, Genetic / methods*
  • Vesiculovirus / genetics

Substances

  • DNA Primers