Gene introduction into granulocyte-macrophage progenitor cells by electroporation: The relationship between introduction efficiency and the proportion of cells in S-phase

Genetic modification of hematopoietic stem cells with nonviral systems: past progress and future prospects

“…83,84 Regarding the requirement for active cell cycling for efficient transfection via electroporation, existing data from cell synchronization experiments are controversial. 80,[85][86][87] Prestimulation of CD34 + cells with recombinant cytokines for various time intervals prior to electroporation has been shown to enhance transfection efficiency and this effect was attributed to the increase in the subpopulation of cells in S phase. 78,83 However, others demonstrated that stimulated and unstimulated CD34 + cells are transfected with equal efficiencies.…”

Optimization of culture conditions to enhance transfection of human CD34+ cells by electroporation

“…Previous studies have demonstrated that electroporation can be a reasonable means of hematopoietic cell gene transfer, and the genes delivered are integrated into the host chromosome. [6][7][8][9][10] Although cell synchronization and electroporation studies have led to controversial conclusions on the requirement for S-phase, [11][12][13] it appears that efficient electroporation of hematopoietic cells requires prestimulation, which results in an apparent increase in the S-phase sub-population. 6,14,15 Ideal culture conditions would promote a high efficiency of gene transfer to ensure not only short-term, but also long-term engraftment and subsequent bone marrow repopulation with each hematopoietic lineage carrying the transgene.…”

Enhanced effect of vascular endothelial growth factor, thrombopoietin peptide agonist, SCF, and Flt3‐L on LTC‐IC and reporter gene transduction from umbilical cord blood CD34+ cells

“…The effect of stromal contact, 2,4 , 20,21 stroma‐conditioned media 22 , or the use of fibronectin 23 and endothelial cell layers 24 are also known to support the growth of early HPCs. Gene transfer into HPCs, for useful clinical manipulation, requires optimal recombinant growth factor combinations to promote stem cell proliferation and gene insertion, while preventing differentiation into committed progenitors 20,25‐27 …”