| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2003: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2002: ¥2,000,000 (Direct Cost: ¥2,000,000)
|
|---|
| Research Abstract |
In this study, we constructed recombinant adeno-associated virus (rAAV) vector expressing enhanced green fluorescent protein (EGEP) under the control of CAG promoter, AV-CAG-EGFP. When the rat common carotid veins were exposed to AV-CAG-EGFP, significant EGFP expression was not detected in the venous specimen. In contrast, we found that human placenta-derived mesenchymal cells (hPDMCs), which reside in placental villi, showed efficient EGFP expression by transduction with AV-CAG-EGFP. After the transduction of AV-CAG-EGFP in the adenoviruses, hPDMCs showed much higher level of EGEP expression than human umbilical vein endothelial cells or rat aortic smooth muscle cells. Moreover, flow cytometric analysis showed discrete positive fraction of EGFP-expressing hPDMCs, which is about 15-20% of the cells infected with AV-CAG-EGFP. Therefore, some cell population in hPDMCs might be highly susceptible to rAAV-mediated gene transduction. In addition, stable EGEP expressions were observed in about 1 % of hPDMCs infected with AV-CAG-EGFP at 4 weeks post-infection. Collectively, hPDMCs have characters favorable for rAAV-mediated gene expression (Microbiol. Immunol. 2003, 47: 109-116). Meanwhile, we found that hPDMCs produce vascular growth factors such as VEGF vigorously. Then, we examined the effect of transplantation of hPDMCs in mouse hindlimb ischemic model, which is supposed to be an animal model of human peripheral vascular disease, arteriosclerosis obliterans. The analysis of vascular ftmction using Laser Doppler and histological analysis showed a favorable outcome by transplantation of hPDMCs in terms of vascular regeneration in this model. Thus, it is possible that rAAV-mediated transduction of therapeutic gene might make transplantation of hPDMCs more beneficial.
|
