Transduction of human vertebrae derived mesenchymal stem cells with GFP lentiviral vectors: assessment of apoptosis and differentiation capacity in vitro

Genetic modification of mesenchymal stem cells (MSCs) may allow further improvement of their therapeutic potential through promotion or suppression of gene expression. An additional benefit of MSCs as a gene delivery system is their lack of immunogenicity, which would confer significant advantage to gene therapy techniques, as the MSC itself may prevent a vector-induced host immune response. Currently lentiviral (LV) transduction is a favorite method to deliver genes into MSCs. The most widely used glycoproteins for pseudotyping LV is the vesicular stomatitis virus glycoprotein (VSV-G), in part due to its very broad tropism and low propensity to cause insertional mutagenesis. However, VSV-G expression has been shown to induce toxicity in certain primary cells including MSCs. Achieving high levels of transgene expression without adversely affecting cell viability and function is crucial if MSCs are to be of benefit using LV based gene modification. In the present study, we investigated LV mediated gene modification of human vertebrae adherent bone-marrow stem cells (vBM-MSC) and assessed in vitro any functional effect of such genetic modification through analyzing viability, apoptosis, surface phenotype and tri-lineage differentiation ability.

Supervisor: Dr. Sreedhar Thirumala
Department: Medical and Molecular Genetics