| Project/Area Number |
60480285
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Hematology
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| Research Institution | Tokyo Women's Medical College |
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Principal Investigator |
MIZOGUCHI HIDEAKI Tokyo Women's Medical College, 医学部, 教授 (70049021)
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| Co-Investigator(Kenkyū-buntansha) |
HOSHINO SHIGERU Tokyo Women's Medical College, 医学部, 助手 (30159131)
KATAHIRA JUNICHI Tokyo Women's Medical College, 医学部, 助手 (00119926)
MOTOJI TOSHIKO Tokyo Women's Medical College, 医学部, 講師 (00101808)
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| Project Period (FY) |
1985 – 1986
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| Project Status |
Completed (Fiscal Year 1986)
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| Budget Amount *help |
¥4,100,000 (Direct Cost: ¥4,100,000)
Fiscal Year 1986: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1985: ¥2,900,000 (Direct Cost: ¥2,900,000)
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| Keywords | interleukin 3 / erythropoietin / megakaryocyte colony-stimulating factor / 巨核球増巾因子 |
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| Research Abstract |
We investigated the effect of purified interleukin 3 (IL-3) and/or purified erythropoietin on murine megakaryocyte colony formation. IL-3 stimulated megakaryocyte colony formation in serum free cultures. These results suggest that IL-3 is a megakaryocyte colony-stimulating factor (MK-CSF). On the other hand, erythropoietin alone did not stimulate megakaryocyte colony formation in serum free cultures. However, when erythropoietin was added to the cultures with IL-3, the number of megakaryocyte colonies and the ploidy values of colony megakaryocytes increased compared to those in the cultures stimulated by IL-3 alone. These results indicate that erythropoietin is not a MK-CSF but a megakaryocyte potenitator (MK-POT).
MK-CSF was purified from a preparation of the urine of patients with aplastic anemia. The eight step purification procedure that includes heat-ethanol treatment, DEAE, SP-Sephadex, Phenyl-Sepharose 6B, <Cu^(++)> chelating-Sepharose 6B, Sephacryl S-200, Sephacryl S-200 rechromatography and DEAE resulted in 20% of the initial biological activity and approximately 100 fold increase in specific activity. This final product showed apparent homogeneity on SDS-PAGE with an estimated molecular weight of 26,000 daltons. Plasma clot culture of mouse bone marrow cells with the purified protein produced a specific activity of 1 x <10^7> solitary megakaryocytes per AU, while IL-3 but not purified MK-CSF supported megakaryocyte colony formation in a serum free culture of mouse bone marrow cells, purified MK-CSF enhanced colony formation supported by IL-3. These results indicate that this purified MK-CSF acts on mouse bone marrow cells as MK-POT. We are planning to investigate the effect of this material on megakaryocyte colony formation by human marrow cells.
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