1998 Fiscal Year Final Research Report Summary
Ex vivo expansion of hematopoietic stem cells using adenovirus
| Project/Area Number |
09671091
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Hematology
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| Research Institution | University of Tokyo |
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Principal Investigator |
CHIBA Shigeru University of Tokyo, Hospital, Intemal Medicine, Associate Professor, 医学部・附属病院, 助手 (60212049)
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| Co-Investigator(Kenkyū-buntansha) |
TAKAHASHI Tokiharu University of Tokyo, Hospital, Intemal Medicine, Clinical Associate, 医学部・附属病院, 医員
OGAWA Seichi University of Tokyo, Hospital, Intemal Medicine, Associate Professor, 医学部・附属病院, 助手 (60292900)
HONDA Hiroaki University of Tokyo, Hospital, Intemal Medicine, Associate Professor, 医学部・附属病院, 助手 (40245064)
MITANI Kinuko University of Tokyo, Hospital, Intemal Medicine, Associate Professor, 医学部・附属病院, 助手 (50251244)
HIRAI Hisamaru University of Tokyo, Hospital, Intemal Medicine, Associate Professor, 医学部・附属病院, 助教授 (90181130)
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| Project Period (FY) |
1997 – 1998
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| Keywords | ex vivo expansion / hematopoietic stem cell / adenovirus vector / epidermal growth factor / CD34+cells / long-term culture-initiating cell / colony-forming unit |
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| Research Abstract |
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-intemalization 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.
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