Open AccessOptimized Lentiviral Transduction of Mouse Bone Marrow-Derived Mesenchymal Stem Cells
Author(s) -
David M. Ricks,
Robert Kutner,
Xian-Yang Zhang,
David A. Welsh,
Jakob Reiser
Publication year - 2008
Publication title -
stem cells and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 114
eISSN - 1557-8534
pISSN - 1547-3287
DOI - 10.1089/scd.2007.0194
Subject(s) - biology , transduction (biophysics) , mesenchymal stem cell , vesicular stomatitis virus , virology , viral vector , stem cell , transgene , ex vivo , genetic enhancement , lentivirus , lymphocytic choriomeningitis , microbiology and biotechnology , cancer research , virus , cytotoxic t cell , in vivo , in vitro , genetics , viral disease , recombinant dna , biochemistry , gene
Mesenchymal stem cells (MSCs) have attracted much attention as potential platforms for transgene delivery and cell-based therapy for human disease. MSCs have the capability to self-renew and retain multipotency after extensive expansion in vitro, making them attractive targets for ex vivo modification and autologous transplantation. Viral vectors, including lentiviral vectors, provide an efficient means for transgene delivery into human MSCs. In contrast, mouse MSCs have proven more difficult to transduce with lentiviral vectors than their human counterparts, and because many studies use mouse models of human disease, an improved method of transduction would facilitate studies using ex vivo-modified mouse MSCs. We have worked toward improving the production of human immunodeficiency virus type 1 (HIV-1)-based lentiviral vectors and optimizing transduction conditions for mouse MSCs using lentivirus vectors pseudotyped with the vesicular stomatitis virus G glycoprotein (VSV-G), the ecotropic murine leukemia virus envelope glycoprotein (MLV-E), and the glycoproteins derived from the Armstrong and WE strains of lymphocytic choriomeningitis virus (LCMV-Arm, LCMV-WE). Mouse MSCs were readily transduced following overnight incubation using a multiplicity of infection of at least 40. Alternatively, mouse MSCs in suspension were readily transduced after a 1-h exposure to lentiviral pseudotypes immediately following trypsin treatment or retrieval from storage in liquid nitrogen. LCMV-WE pseudotypes resulted in efficient transduction of mouse MSCs with less toxicity than VSV-G pseudotypes. In conclusion, our improved production and transduction conditions for lentiviral vectors resulted in efficient transduction of mouse MSCs, and these improvements should facilitate the application of such cells in the context of mouse models of human disease.