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A Potential Molecular Approach to Ex Vivo Hematopoietic Expansion
With Recombinant Epidermal Growth Factor Receptor-Expressing Adenovirus
Vector
Tokiharu Takahashi,
Kaoru Yamada,
Tomoyuki Tanaka,
Keiki Kumano,
Mineo Kurokawa,
Tsuyoshi Takahashi,
Naoto Hirano,
Hiroaki Honda,
Shigeru Chiba,
Kohichiro Tsuji,
Yoshio Yazaki,
Tatsutoshi Nakahata, and
Hisamaru Hirai
From The Third Department of Internal Medicine, the Department of
Transfusion and Immunohematology, and the Department of Cell Therapy & Transplantation Medicine, Faculty of Medicine, University of Tokyo,
Bunkyo-ku, Tokyo, Japan; and the Department of Clinical Oncology,
Institute of Medical Science, University of Tokyo, Minato-ku, Tokyo,
Japan.
Ex vivo expansion of hematopoietic stem cell (HSC) is an attractive
technology for its potency of a variety of clinical applications. Such
a technology has been achieved to some extent with combinations of
various cytokines or continuous perfusion cultures. However, much more
improvement is required especially for expansion of primitive
hematopoietic progenitors. We propose here a novel molecular approach
that might have the potential to compensate the current expansion. We designed an adenovirus vector to transiently express human epidermal growth factor receptor (EGFR), which is known to
transduce only a mitogenic, but not a differentiation signal to mouse
bone marrow cells on human purified CD34+ peripheral
blood (PB) cells, and tried to expand these cells with EGF ex vivo.
Because we found that exposure of CD34+ PB cells to
cytokines induced surface expression of adenovirus-internalization receptor and rendered these cells permissive to adenovirus infection, we infected these cells with the adenovirus vector carrying EGFR gene
in the presence of cytokines. Two-color flow cytometric analysis demonstrated that 60.3% ± 22.4% of CD34+ cells
expressed the adenovirus-mediated EGFR. Moreover, long-term culture-initiating cell assay showed that adenovirus
vector could transduce more primitive progenitors. Subsequently, we
tried to expand these cells in suspension culture with EGF for 5 days. Methylcellulose clonal assay showed that EGF induced 5.0- ± 2.4-fold proliferation of the colony-forming unit pool during 5 days of expansion. The simple procedure of efficient adenovirus gene delivery to immature hematopoietic cells proved promising, and this technique was potentially applicable for a novel strategy aiming at ex vivo expansion of hematopoietic progenitors.
Blood, Vol. 91 No. 12 (June 15), 1998:
pp. 4509-4515
© 1998 by The American Society of Hematology.
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