1991 Fiscal Year Final Research Report Summary
The mechanisms of proliferation and differentiation of hematopoietic multipotent stem cells.
| Project/Area Number |
01570542
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Pediatrics
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| Research Institution | Yokohama City University, School of Medicine |
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Principal Investigator |
SASAKI Hideki Yokohama City Univ. School of Medicine, Pediatrics, Associate Professor, 医学部, 助教授 (50106316)
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| Co-Investigator(Kenkyū-buntansha) |
INOUE Tohru Yokohama City Univ. School of Medicine, Pathology, Associate Professor, 医学部, 助教授 (50100110)
MATSUYAMA Shusuke Yokohama City Univ. School of Medicine, Pediatrics, Professor, 医学部, 教授 (20045983)
IKUTA Koichiro Yokohama City Univ. School of Medicine, Pediatrics, Assistant Professor, 医学部, 講師 (80159590)
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| Project Period (FY) |
1989 – 1991
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| Keywords | Hemopoietic Stem Cells / Interleukin / Interleukin 6 / Stem Cell Factor / Leukemia Inhibitory Factor / Erythropoietin |
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| Research Abstract |
To clarify tlie mechanisms of proliferation and differentiation of multipotent hemopoietic stem cells, we investigated the effects of IL-3, IL-6, SCF, LIF and EPO on early appearing CFU-S(CFU-S_8)and late appearing CFU-S(CFU-S_<12>)in short-term suspension culture. We also employed the ^3H-TdR suicide method to test whether the modulation of CFU-S by IL-3 and EPO involved the proliferative process. A single addition of IL-3 or SCF increased the numbers of both subpopulation of CFU-S recovered in culture with dose-dependent fashion, while EPO, at least up to 10u/ml, increased solely CFU-S_8. Further, dose-response curves of CFU-S_8 and CFU-S_<12> to IL-3 and SCF were different, which confirmed the speculation that CFU-S_8 and CFU-S_<12> are relatively different subpopulation. Our results including ^3H-TdR suicide suggested that EPO acted on both of CFU-S_8 and CFU-S_<12> to stimulate proliferation with differentiation into more mature population in erythroi. d lineage, and that IL-3 acted not only on these two subpopulation but also on more primitive stem cell population which was resistant to 5FU. The investigation on ihe action of SCF in detail is on going. Another two cytokines, IL-6 and LIF, showed synergistic stimulating activity with IL--3. The physiological roles of these factors in the regulatory mechanisms of stem cell proliferation are still unknown. However, studies of tlie action of these cytokines on hemopoietic stem cells are important research fields in regarding to clinical uses such as in votro expansion for transplantation.
Human BFU-E, CFU-GM, and CFIJ-Mix were 20x enriched by one-step immunomagnetic beads separation using anti-CD_<34> antibody. Basical studies suggested multi-step separation procedures should be required for further improvement of the efficiency for clinical use.
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