Historically, efficient transduction of hematopoietic stem cells (HSC) to study the role of specific genes on HSC function, as well as to broaden the potential of gene therapy for hematopoietic related diseases has relied on our ability to design vectors capable of delivering the gene of interest without affecting HSC function. While retroviruses have been used extensively for this purpose, HIV-derived lentiviruses prove superior for transduction of quiescent HSC due to their ability to infect nondividing cells. The design of the vector and the quality of the lentiviral preparation are the key elements to obtain reproducible consistent results that will eventually be translated into the clinic. This chapter describes the preparation of concentrated lentiviruses and the transduction of HSC to obtain long-term engraftment with persistent gene transfer and expression of the desired transgene.
Keywords: Hematopoietic stem cells; Lentiviral transduction; Third-generation lentiviruses; Transfection; Ultracentrifugation.