Project/Area Number |
12670972
|
Research Category |
Grant-in-Aid for Scientific Research (C)
|
Allocation Type | Single-year Grants |
Section | 一般 |
Research Field |
Hematology
|
Research Institution | University of Tsukuba |
Principal Investigator |
NAGASAWA Toshiro Institute of Clinical Medicine, University of Tsukuba, Professor, 臨床医学系, 教授 (70014298)
|
Co-Investigator(Kenkyū-buntansha) |
ISHIDA Yoji Division of Hematology, Iwate Medical School, Assistant Professor, 医学部, 講師 (70151389)
NAKAUCHI Hiromitsu Institute of Basic Medicine, University of Tsukuba, Professor, 基礎医学系, 教授 (40175485)
|
Project Period (FY) |
2000 – 2001
|
Project Status |
Completed (Fiscal Year 2001)
|
Budget Amount *help |
¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 2001: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2000: ¥1,600,000 (Direct Cost: ¥1,600,000)
|
Keywords | megakaryopoiesis / in vitro induced platelets / platelet transfusion |
Research Abstract |
The final goal of such megakaryopoiesis is the platelet production. However, it is still difficult to recover a large number of platelets from a culture system. In this study, it is aimed to recover in vitro induced platelets from purified hematopoietic stem cells. One hundred murine early pluripotent stem cells (EPSC) (CD34^-/c-kit^+/Sca-1^+/Lin^-) sorted by FACS vantage were co-cultured on murine stromal cell line (HESS-5) in the presence of stem cell factor (10 ng/ml), IL-3 (100 ng/ml), IL-6 (100 ng/ml), sIL-6r (100 ng/ml) and Tpo (20 ng/ml) for 7-10 days. 100 early pluripotent stem cells produced about 6000 megakaryocytes and 4-5 x 10^5 plateletes in the supernatant by the combination of SCF + IL-3 + Tpo + IL-6 + sIL-6r + SCF + IL-3. The maximum thrombin-induced aggregation of in vitro induced-platelets was about 50 % of the control. The in vivo recovery of infused platelets was 75 % of the control when the platelets were infused in the irradiated thrombocytopenic mice. In vitro induced platelets derived from murine hematopoietic stem cells have proved effective for in vivo hemostasis.
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