title: Efficient transduction of equine adipose-derived mesenchymal stem cells by VSV-G pseudotyped lentiviral vectors.
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.
authors.affiliation: School of Biomedical Sciences, Charles Sturt University, Wagga Wagga, NSW, Australia.
authors.forename: Gayle F
authors.initials: GF
authors.lastname: Petersen
authors.affiliation: School of Animal and Veterinary Sciences, Charles Sturt University, Wagga Wagga, NSW, Australia.
authors.forename: Bryan
authors.initials: B
authors.lastname: Hilbert
authors.affiliation: School of Animal and Veterinary Sciences, Charles Sturt University, Wagga Wagga, NSW, Australia.
authors.forename: Gareth
authors.initials: G
authors.lastname: Trope
authors.affiliation: School of Biomedical Sciences, Charles Sturt University, Wagga Wagga, NSW, Australia.
authors.forename: Wouter
authors.initials: W
authors.lastname: Kalle
authors.affiliation: School of Biomedical Sciences, Charles Sturt University, Wagga Wagga, NSW, Australia. Electronic address: pstrappe@csu.edu.au.
authors.forename: Padraig
authors.initials: P
authors.lastname: Strappe
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D000704:Analysis of Variance; D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002454:Cell Differentiation / Q000502:physiology; D017931:DNA Primers / Q000235:genetics; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D006734:Horse Diseases / Q000235:genetics / Q000628:therapy*; D006736:Horses; D016086:Lentivirus / Q000235:genetics*; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D057054:Molecular Imaging / Q000662:veterinary; D014161:Transduction, Genetic / Q000379:methods*; D018116:Vesiculovirus / Q000235:genetics
chemical_list: D017931:DNA Primers
keywords: Adipose; Equine; Gene delivery; Green fluorescent protein; Lentiviral vector; Mesenchymal stem cells