Stroma-conditioned medium and sufficient prestimulation improve fibronectin fragment-mediated retroviral gene transfer into human primitive mobilized peripheral blood stem cells through effects on their recovery and transduction efficiency

Leukemia
D A BreemsR E Ploemacher

Abstract

Mobilized peripheral blood stem cells (PBSC) are an attractive vehicle for cancer gene therapy. However these stem cells may have a reduced proliferative capacity due to previous cytotoxic chemotherapy treatment of the patient. In addition, primitive hematopoietic stem cells (HSC) from mobilized peripheral blood are almost exclusively quiescent, which makes it hard to induce proliferation in vitro and thus to improve stable transduction of introduced genes into a sufficiently large number of primitive stem cells. In this study CD34-selected mobilized PBSC from lymphoma and myeloma patients were used as target cells for retroviral-mediated gene transfer using a clinically relevant cell- and serum-free supernatant transduction protocol. We have investigated various parameters that may contribute to an improvement of the poor transduction efficiency of the primitive HSC, including prestimulation time, the use of the carboxy-terminal fibronectin fragment CH-296, as well as stromal cell line conditioned media. Retroviral supernatant transduction in combination with CH-296 increased significantly the gene transfer efficiency as compared to supernatant alone and made the use of polycations redundant. Gene transfer of primitive HSC (co...Continue Reading

Citations

May 7, 1999·Cancer radiothérapie : journal de la Société française de radiothérapie oncologique·B DubrayJ M Cosset
Apr 2, 2005·Methods in Cell Science : an Official Journal of the Society for in Vitro Biology·Dimitrios G Bouzianas
Oct 30, 2001·Leukemia·M Bhatia
Jun 6, 2006·Laboratory Hematology : Official Publication of the International Society for Laboratory Hematology·Azza A MohamedRaafat M Riad

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