Retroviral transduction of quiescent haematopoietic cells using a packaging cell line expressing the membrane-bound form of stem cell factor

Gene therapy vectors based on murine retroviruses are unable to transduce n on-dividing cells. This has proven a particular problem in the haematopoiet ic system where the target cells of choice, the pluripotent stem cells are quiescent. In an attempt to circumvent this difficulty we have constructed a retroviral producer line that expresses the membrane bound form of human recombinant stem cell factor (SCF) on its cell surface. This should enable the retroviral producers to deliver a growth signal to the target cells sim ultaneous with their exposure to retrovirus. We tested the ability of these modified producers to transduce a growth factor-starved SCF-dependent cell line (TF-1) and demonstrated that these cells, though quiescent, can still be successfully transduced This approach was extended to targeting of umbi lical cord blood CD34(+) cells, a predominantly quiescent population that n ormally require the addition of cytokines for efficient transduction. Using the SCF-expressing producer line in the absence of exogenously added cytok ines, we observed a marked stimulation in transduction efficiency over that achieved using the parent producer line alone. Colonies derived from these cells arising in semi-solid media were also shown to be positive for expre ssion of a retrovirally encoded repenter gene.