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- W4242959092 abstract "307 Both viral and non-viral vectors have been quite limited in their ability to accomplish highly efficient gene delivery to relevant target cellsin vivo. As an alternative vector strategy, parenchymal cells have been employed. In this approach, cells are removed from the body and genes are transferred into the cells ex vivo, followed by reimplantation. Thus, the transduced cell becomes the ultimate vector for gene delivery. The role of circulating CD34+ cells as endothelial progenitors, or angioblasts, has recently been described. We sought to develop a novel cellular vector for gene therapy that may allow the delivery of therapeutic molecules into foci of angiogenesis. METHODS: We isolated CD34+ cells from rhesus monkeys. Bone marrow was obtained after mobilization with G-CSF, and mononuclear cells were isolated by Ficoll sedimentation. CD34+ cells were then purified by column immunoabsorption. Cells were maintained in plastic wells in culture medium enriched with IL-3, IL-6, and SCF at 37C in a humidified, 5% CO2 atmosphere. Cells were transduced ex vivo with TOZ.1, a highly efficient (>99%) and nontoxic herpes vector encoding both the reporter gene. LacZ and thymidine kinase. Infection was done with a multiplicity of infection (MOI) of 3 infectious particles per cell for 12 hours. To evaluate the localization capacity of CD34+ cells into areas of angiogenesis, we established an in vivo model based in skin autografts. In addition, we implanted s.c, in a separate region, Matrigel and Gelfoam impregnated with the vascular endothelial growth factors VEGF and basic FGF. Transduced cells were infused intravenously two days after graft and pellet implantation. Biopsies were obtained periodically from the graft and pellets. Transduced cells expressing LacZ-encodedβ-galactosidase were revealed by X-gal staining, and an antibody against factor VIII was used as endothelial marker. As a further means to show the expression of genes delivered by CD34+ cells at the angiogenesis areas, treatment with ganciclovir (GCV) was given (5 mg/kg i.v. for 14 days) 15 days after CD34+ cell infusion. RESULTS: Both in the skin graft and implants, numerous small, round, intensely blue cells were identified in neovascular structures stained by factor VIII. After 10 days of treatment with GCV, the subcutaneous pellets disappeared, suggesting conversion of GCV to its toxic metabolites in the angiogenesis sites. Animals nontreated with GCV, in contrast, kept palpable implants during the study time. Importantly, no symptom or sign of toxicity has been observed in three different primates infused with TOZ.1-transduced CD34+ cells. CONCLUSIONS: CD34+ cells can indeed localize into areas of angiogenesis and express reporter and toxin genes therein. This allows evaluating their potential as cellular vehicles with capacity for localization and gene delivery into areas of angiogenesis, including graft implants." @default.
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- W4242959092 date "1998-06-01" @default.
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- W4242959092 title "GENETICALLY MODIFIED CD34+ CELLS AS VECTORS FOR IN VIVO GENE DELIVERY TO AREAS OF ANGIOGENESIS" @default.
- W4242959092 doi "https://doi.org/10.1097/00007890-199806270-00326" @default.
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