1992 Fiscal Year Final Research Report Summary
Cell culture and application of fish cells with defferentiated functions
| Project/Area Number |
03660202
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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 |
General fisheries
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| Research Institution | NIHON UNIVERSITY |
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Principal Investigator |
WATANABE Tasuku Ph.D., Nihon University, Department of Agriculture and Veterinary Medicine, Associate Professor, 農獣医学部, 助教授 (30095517)
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| Project Period (FY) |
1991 – 1992
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| Keywords | Liver cells / Glycoprotein / Yamame / Hematopoietic cell / Stem cell / Semi-solid agar / Oncogene |
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| Research Abstract |
Yamame embryonal liver cell line, YEL-13, produces and releases large glycoprotein, which was stained with Giemsa and PAS. Electron microscopically, the cells had polymorphic unclei and well developed microfibril, and lost the morphological properties of liver parenchymal cells. Electron-dense graunles, surrounded by cell membrane, budded and were released from the surface of cells. Glycoprotein, isotlated from the conditioned medium of YEL-13, were purified partially with Parcoll zonal density gradient centrifugation and had the density of 1.04g/ml. Antiserum to this fraction stained the glycoprotein of YEL-13 and liver parenchymal cells of Yamame, strongly. These results suggest that YEL-13 became to produce these glycoprotein super-fluously with the immortalization in in vitro. We started the studies on the role of the glycoprotein in liver and on the analysis of sugar sequences using HPLC.
The hematopoietic stem cells of carp can multiply to from many colonies in semi-solid agar media when supplemented with carp serum. The colony forming cells varied from undifferentiated stem cells to differentiated granulocytes, neutrophils, baso-phils, and monocytes. When used the media supplemented with the serum of adjuvant-injected carp, more colonies were formed than with normal carp serum, As this hematopoietic stem cell culture system of carp is available for the investigation of fish colony stimulate factors (CSF), we started the purification studies of CFSs from carp sera. We also started the oncogene transfection study to carp hematopoietic cells for the immortilization of white blood cells.
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