| Project/Area Number |
17500294
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Laboratory animal science
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| Research Institution | Junior College of Tokyo University of Agriculture |
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Principal Investigator |
KADA Hidemi Tokyo University of Agriculture, Department of Bioproduction Technology, Assistant Professor, 生物生産技術学科, 講師 (40214340)
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| Co-Investigator(Kenkyū-buntansha) |
SATO Kahei Nihon University, Department of applied Biological Science, Professor, 生物資源科学部, 教授 (20059778)
WAKUI Kenji Tokyo University of Agriculture, Department of Bioproduction Technology, Assistant Professor, 生物生産技術学科, 講師 (40331433)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 2006: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2005: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | oocyte / culture in vitro / growth in vitro / maturation in vitro / oocyte maturation / co-culture / mouse / oocyte-granulosa cell complexes / 生殖細胞 |
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| Research Abstract |
The aim of this study was to establish the efficient culture method for mouse oocytes in vitro. A novel system was designed by co- culture with feeder cells derived from ovaries. B6CBF1 female mice (12 days old) were used in this study. Oocyte-granulosa cell complexes (OGCs) were isolated from ovaries by collagenase digestion in Leibovi's L-15 medium with 5% fetal bovine serum (FBS). The OGCs were group-cultured for 10 or 12 days on the feeder cells on collagen-coated insert membrane in a-minimum essential medium (MEM) supplemented with 100mlU/ml follicle-stimulating hormone, 10m1U/mIluteinizing hormone, 50ng/ml insulin-like growth factor type I, 50ng/ml stem cell factor at 37℃. Half amount of medium was exchanged on alternate days. The oocyte growth in vitro was achieved by this system. The oocyte maturation was induced by addition of final concentration of 1.5lU/ml human chorionic gonadotrophin and 5ng/ml epidermal growth factor on day 10 or 12 of culture. Release and mucification of the OCCs was observed at 17 hours after the maturation induction. The survival rate, the volume increase, the maturation competence and the developmental competence were improved by addition of gonadotropin, growth factor and co-culture, compared with old system. In culture in vitro from fetal ovaries (16-17dpc), the IVG-IVM oocytes showed no embryonic development, unfortunately. Thus, further improvement of culture conditions is required. However, the effectiveness of this culture system was shown in culture in vitro from the OGCs.
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