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This article presents our studies on the adenoviral transduction efficiency, level of transgene expression, cell cycle status, and multilineage reconstitution ability of human CD34+ hematopoietic cells transduced under proliferating and survival growth conditions. Bone marrow and umbilical cord blood CD34+ cells were cultured in serum-free medium under survival conditions with thrombopoietin (Tpo) alone, or under proliferating conditions with Tpo, c-Kit ligand (KL), and Flt3 ligand (FL). Adenoviral vectors carrying the enhanced green fluorescent protein (EGFP) gene under the control of the PGK-1 promoter were used to transduce CD34+ cells. Approximately 10% of CD34+ cells were EGFP+ under both culture conditions. In contrast, up to 50% of CD34+CD38- cells were EGFP+, whereas a maximum of 8% of CD34+CD38(high) cells were EGFP+ (p < 0.001). Both colony-forming unit cells (CFU-C) and 5-week long-term culture-initiating cells (LTC-ICs) were efficiently transduced. Under survival conditions, a substantial fraction of transduced CD34+ cells remained quiescent. The nondividing CD34+EGFP+ cells contained LTC-ICs capable of reconstituting longterm culture for as long as 10 weeks. CD34+EGFP+ cells also retained the ability to engraft and multilineage-reconstitute NOD/SCID mice. These observations demonstrate that primitive human hematopoietic progenitor cells can be efficiently transduced by adenoviral vectors.

Original publication




Journal article


Hum Gene Ther

Publication Date





1313 - 1327


ADP-ribosyl Cyclase, ADP-ribosyl Cyclase 1, Adenoviridae, Animals, Antigens, CD, Antigens, CD34, Antigens, Differentiation, Bone Marrow Cells, Cell Division, Cell Lineage, Cell Survival, Fetal Blood, Gene Expression, Genetic Vectors, Green Fluorescent Proteins, Hematopoietic Stem Cell Transplantation, Hematopoietic Stem Cells, Humans, Luminescent Proteins, Membrane Glycoproteins, Membrane Proteins, Mice, Mice, SCID, NAD+ Nucleosidase, Stem Cell Factor, Thrombopoietin, Transduction, Genetic