| Project/Area Number |
11460132
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| Research Category |
Grant-in-Aid for Scientific Research (B).
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Applied animal science
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| Research Institution | AZABU UNIVERSITY |
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Principal Investigator |
TACHI Chikashi Azabu University, School of Veterinary Madicine, Professor., 獣医学部, 教授 (30011711)
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| Co-Investigator(Kenkyū-buntansha) |
YOKOYAMA Minesuke Mitsubishi-Kasei Institute of Life Sciences, Center for Mouse Genomic Engineering, Head., 生殖工学開発室, 室長
TAKAGI Nobuo Hokkaido University, Graduate School for Environmental Eearth Science, Professor., 大学院・地球環境科学研究科, 教授 (20001852)
KASHIWAZAKI Naomi Azabu University, School of Veterinary Madicine, Lecturer., 獣医学部, 講師 (90298232)
HISAMATSU Shin Azabu University, Faculty of Environmental Health, Lecturer., 環境保健学部, 講師 (10198997)
SAWASAKI Toru University of Tokyo, Graduate School of Agriculture and Life Sciences, Professor., 大学院・農学生命科学研究科, 教授 (00012047)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥14,700,000 (Direct Cost: ¥14,700,000)
Fiscal Year 2000: ¥5,800,000 (Direct Cost: ¥5,800,000)
Fiscal Year 1999: ¥8,900,000 (Direct Cost: ¥8,900,000)
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| Keywords | ES cells / PGC / Germ line cells / SCF / Feeder cells / 4C9 / 2C9 / Goats / EG細胞 / 始原生殖細胞 / 4C9抗体 / 2C9抗体 / 卵黄嚢細胞 / ヤギ / ブタ |
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| Research Abstract |
In artyodactyls, the production of transgenic animals or clonal animals by means of nuclear transfer from somatic cells, is rapidly becoming practical means in animal industries despite many unsolved problems. One of such problems concerns with the availability of unfertilized ova which are often difficult to obtain in sufficient quantity. If ES/EG cells with the XX karyotype could be induced to enter into the germ line in vitro, and ultimately to differentiate into ova, an important progress will be made in various fields of animal biotechnology, especially those dealing with large mammals. The purpose of present studies is to carry out basic research exploring the possibility of developing techniques which enable the convrsion of ES/EG cells to the germ line. In the following, the results of experiments are summarized. 1) Immunocytochemical studies of a murine ES cell line revealed the presence of cells which reacted with 4C9 (monoclonal antibody against a PGC marker protein) and 2C9
… More
(monoclonal antibody raised against chicken PGC) positively, probably respresenting germ line cells of the population. Culture of 4C9/2C9 positive ES cells under the conditions which stimulated PGC proliferation did not cause significant stimulation of these cell. Analysis of gene structure and functions of vasa which is knoown to be expressed in PGC was made. 2) In order to establslish an in vitro culture system for caprine PGC, as a model for bovidae animals in general, caprine SCF (synonyms : KL and MGF) cDNA was cloned, and usd for the establishement of the feeder cell lines where the membrane-bound as well as soluble form of cSCF are expressed. 3) ES cells with XX karyotype were analyzed for the expression of Xist and its antisense gene Tsix both of which play crucail roles in Lyonization of X chromosomes. The expression of the 2 genes was confirmed. 4) The IVM and IVF procedures for porcine oocytes were investigated. Basic studies were done for the application of ribozymes in the genetic manipulation of farm animals. 5) Summary. In recent years, rapid progress has been made in the studies on conversion of ES/EG cells to the germ line. The present research project yielded valuable clues to approach the problem. Less
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