256. Novel Therapeutic Strategy for Ischemic Stroke by Bone Marrow Stromal Cells and Ex Vivo HGF and FGF2 Genes Transfer with Replication-Incompetent HSV-1 Vector

The bone marrow stromal cells (MSCs), including the primitive pluriopotent mesenchymal stem cells (PMSCs) and the multipotent adult progenitor cells (MAPCs), are attractive targets for cell and gene therapy for various central nervous system disorders.Material and method: For gene modification of MSCs, we used a replication-incompetent HSV-1 vector "1764/4-", triple-deletion mutant of the ICP4, ICP34.5, and VP16. This vector has very high ability of gene transduction to MSC, 70-75% at MOI 10 and 95% at MOI 50, and its expression can carry on at least one month at high level. Two kinds of vectors expressing HGF and FGF-2 were prepared for this study. FGF-2 vector contains the signal sequence of Interleukin-2 (IL-2ss) ahead of FGF-2 gene to enhance its extracellular secretion, because native FGF2 gene don't have the excretion signal. Native MSCs, HGF-MSCs, IL2ss+FGF-2-MSCs and PBS were administered directly into the lesioned brain 24 hours after transient middle cerebral artery occlusion (MCAO) of adult Wister rats.Results: At day 14, each gene modified MSC group showed significant improvement in modified neurological severity score (mNSS) and apparent decreasing of infarct volume compared with native MSC or sham-operated group. (p