Gene Marking : Retroviral Vector-Mediated Gene Transfer into CD34-Enriched Human Bone Marrow Stem Cells (Gene Therapy : Advances in Research and Treatment)

In order to improve the efficiency of gene transduction into human hematopoietic stem cells by retrovirus vectors, we conducted an in vitro study to determine the optimal conditions. Cells used as the target were CD34-enriched human bone marrow stem cells. Retrovirus vectors used were LNL6 and G1Na40, both carrying the Neomycin-resistant (Neo-R) gene as a genetic marker. Transduction efficiency varied from 5.1% to 35.0%, depending on the factors and procedures in the experiments, which cornprised MOI, the duration of exposure of cells to the vectors, and conduct/nonconduct of centrifuge of the cells during the exposure. The most effective gene transduction into CFV-GM was found to be a 48h-liquid culture with growth factors followed by 48h-vector exposure with the spin transduction