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.
Copyright © 2014 Elsevier Ltd. All rights reserved.
Publication Date: 2014-10-06 PubMed ID: 25443656DOI: 10.1016/j.rvsc.2014.09.004Google Scholar: Lookup
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- Comparative Study
- Evaluation Study
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This study analyses the utilization of lentiviral vectors for gene modification in equine adipose-derived mesenchymal stem cells (EADMSC), with the goal of improving the cells’ therapeutic capabilities for treating horse musculoskeletal injuries.
Research Context
- The research is centered on enhancing the capabilities of equine adipose-derived mesenchymal stem cells (EADMSC), known for their unique ability to help treat various equine musculoskeletal ailments. This is achieved either through the regeneration of diseased or damaged tissue or secretion of immunomodulatory molecules.
- Lentiviral vectors are the primary technology in focus to boost these capabilities as they present a reliable and efficient means of gene delivery to these cells.
Methodology
- Two types of GFP (Green Fluorescent Protein) expressing lentiviral vectors were used: one pseudotyped with the G glycoprotein from the vesicular stomatitis virus (termed V-GFP) and the other one pseudotyped with the baculovirus gp64 envelope protein (termed G-GFP). This is the first time that the G-GFP lentiviral vector has been used in such a study.
- The general research approach involved producing high titre V-GFP and G-GFP lentiviral vectors and administering them to EADMSC. Transduction efficiency was then analyzed through flow cytometry.
Findings
- The V-GFP lentiviral vector showed impressive transduction efficiency, while the G-GFP vector presented poor transduction capabilities for EADMSC.
- Importantly, the process did not drastically impact cell viability or doubling time. An insignificant effect on the chondrogenic differentiation capability of the EADMSC was also observed.
- Transduction led to the prolonged expression of GFP in the EADMSC, spread over at least four cell passages.
Conclusion
- The findings of the research suggest that lentiviral vectors, particularly those pseudotyped with V-GFP, are potentially viable tools for gene delivery into EADMSC. This could help to augment their therapeutic efficacy in treating equine musculoskeletal injuries.
Cite This Article
APA
Petersen GF, Hilbert B, Trope G, Kalle W, Strappe P.
(2014).
Efficient transduction of equine adipose-derived mesenchymal stem cells by VSV-G pseudotyped lentiviral vectors.
Res Vet Sci, 97(3), 616-622.
https://doi.org/10.1016/j.rvsc.2014.09.004 Publication
Researcher Affiliations
- School of Biomedical Sciences, Charles Sturt University, Wagga Wagga, NSW, Australia.
- School of Animal and Veterinary Sciences, Charles Sturt University, Wagga Wagga, NSW, Australia.
- School of Animal and Veterinary Sciences, Charles Sturt University, Wagga Wagga, NSW, Australia.
- School of Biomedical Sciences, Charles Sturt University, Wagga Wagga, NSW, Australia.
- School of Biomedical Sciences, Charles Sturt University, Wagga Wagga, NSW, Australia. Electronic address: pstrappe@csu.edu.au.
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
Citations
This article has been cited 7 times.- Barrachina L, Ivanovska A, Eslami Arshaghi T, O'Brien A, Cequier A, Murphy M, Hollinshead F, Rodellar C, Barry F. Generation of equine induced pluripotent stem cells from cells of embryonic, perinatal and adult tissues. Stem Cell Res Ther 2025 Oct 8;16(1):547.
- Arduini A, Katiyar H, Liang C. Progress in Pseudotyping Lentiviral Vectors Towards Cell-Specific Gene Delivery In Vivo. Viruses 2025 May 31;17(6).
- Deng L, Liang P, Cui H. Pseudotyped lentiviral vectors: Ready for translation into targeted cancer gene therapy?. Genes Dis 2023 Sep;10(5):1937-1955.
- Thampi P, Samulski RJ, Grieger JC, Phillips JN, McIlwraith CW, Goodrich LR. Gene therapy approaches for equine osteoarthritis. Front Vet Sci 2022;9:962898.
- Mançanares ACF, Cabezas J, Manríquez J, de Oliveira VC, Wong Alvaro YS, Rojas D, Navarrete Aguirre F, Rodriguez-Alvarez L, Castro FO. Edition of Prostaglandin E2 Receptors EP2 and EP4 by CRISPR/Cas9 Technology in Equine Adipose Mesenchymal Stem Cells. Animals (Basel) 2020 Jun 23;10(6).
- Winter RL, Seeto WJ, Tian Y, Caldwell FJ, Lipke EA, Wooldridge AA. Growth and function of equine endothelial colony forming cells labeled with semiconductor quantum dots. BMC Vet Res 2018 Aug 23;14(1):247.
- Bandara N, Gurusinghe S, Lim SY, Chen H, Chen S, Wang D, Hilbert B, Wang LX, Strappe P. Molecular control of nitric oxide synthesis through eNOS and caveolin-1 interaction regulates osteogenic differentiation of adipose-derived stem cells by modulation of Wnt/β-catenin signaling. Stem Cell Res Ther 2016 Dec 7;7(1):182.
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