| Project/Area Number |
06671109
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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 |
Hematology
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| Research Institution | DOKKYO UNIVERSITY |
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Principal Investigator |
FURUKAWA Toshiharu DOKKYO UNIVERSITY,SCHOOL OF MEDICINE PROFESSOR, 医学部, 教授 (70049154)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1995: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1994: ¥900,000 (Direct Cost: ¥900,000)
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| Keywords | Eosinophil / Videomicrography / Compression / Generation time / IL-3 / IL-5 / Motility / Human cord blood / 顕微鏡ビデオ撮影 / 核分葉 / 分化 / 運動パターン / rhIL-3 / rhIL-5 / 長期顕微鏡ビデオ / 増殖・分化 / 前駆細胞 |
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| Research Abstract |
In the present study, we used a new device to compress eosinophil precursors allowing their fine structures to become more visible without losing their ability to proliferate and differentiate. Mononuclear cells separated from cord blood were cultured in Iscove's modified Dulbecco medium containing 10% bovine fetal serum, rhIL-3 and rhIL-5, for 9 days in a humidified atmosphere containing 5% CO_2. Non-adhering cells were harvested and placed in a human fibrin clot. A small cover slide was placed over the specimen. The cells were compressed using a new device with a forked rod applied on the cover slide, the pressure applied was adjusted by a screw. The cells were cultured in the same medium as described above. A Nikon phase-contrast inverted microscope placed in a thermostatic chamber was used to obtain time-lapsed video-micrography, over 5-9 days of culture. During this observation period, most of the eosinophil precursors divided 2-3 times to yield freely moving mature eosinophils. Generation time varied from 17 hours 8 minutes to 96 hours 3 minutes. A tendency towards a longer generation time in more mature eosinophils was observed. Pattern of motility in the eosinophils changed along with their differentiation : the most immature eosinophil precursors with a round or oval nucleus showed little motility except for occasionally throwing a wide veil ; intermediately mature eosinophil precursors repeatedly prodded and retracted a reatively narrow pseudopod in one direction ; mature eosinophils showed relatively fast motion taking a shape of a broad rod with a round posterior end.
This study showed that human eosinophil precursors could continue to proliferate and differentiate under appropriate compression which allow their fine strucutral changes to be more easily visualizad.
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