| Project/Area Number |
60870059
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| Research Category |
Grant-in-Aid for Developmental Scientific Research
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| Allocation Type | Single-year Grants |
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| Research Field |
Obstetrics and gynecology
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| Research Institution | KEIO UNIVERISTY |
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Principal Investigator |
RIHACHI IIZUKA Keio Univeristy, Shool of Medicine, Dept. of ob/Gyn, 医学部産婦人科学教室, 教授 (70050995)
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| Co-Investigator(Kenkyū-buntansha) |
久慈 直昭 慶応義塾大学, 医学部産婦人科学教室, 助手 (80169987)
MICHIYA NATORI Keio University, School of Medicine, Dept. of ob/Gyn, 医学部産婦人科学教室, 講師 (80101913)
TSUNEHISA MAKINO Keio University, School of Medicine, Dept. of ob/Gyn, 医学部産婦人科学教室, 講師 (30085758)
MASATU MORISADA Keio University, School of Medicine, Dept. of ob/Gyn, 医学部産婦人科学教室, 講師 (40051552)
TOSHIFUMI KOBAYASHI Keio Unversity, School of Medicine, Dept. of ob/Gyn, 医学部産婦人科学教室, 助教授 (30051460)
KIYOO TAsNABE Keio Univerity, Shool of Medicine, Dept. of ob/Gyn
末岡 浩 慶応義塾大学, 医学部産婦人科学教室, 助手 (90162833)
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| Project Period (FY) |
1985 – 1987
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| Project Status |
Completed (Fiscal Year 1987)
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| Budget Amount *help |
¥9,000,000 (Direct Cost: ¥9,000,000)
Fiscal Year 1987: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1986: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1985: ¥7,000,000 (Direct Cost: ¥7,000,000)
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| Keywords | Cryopreservation / Human embryo / Human oocyte / Electron microscopic exmaniation / 体外受精-胚移植 / 凍結保存 / 未受精卵 / 2細胞期 / マウス / 2段階融解法 / 電子顕微鏡 / グルコース / 体外受精 / 凍結保護剤 / プログラムフリーザー / マイクロマニピュレーター |
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| Research Abstract |
Cryopreservation of embryos and oocytes has been widely performed in the field of reproductive science to study on safety, cryprotectants, freezing methods, etc. In this report, results of ceyopresercation of murine and human embryos and oocytes were pressnted to discuss safer mothods of freezing and thawing, length of storage period, ultrastructural changes in frozen-thawed oocytes, and a future prespective of human embryo cryopreservation in in vitro fertilization and embryo transfer. Combination of slow cooling ( at a rate of 0.3 dagress Centigrade per minute to minus 80 degrees C.) and slow thawing yielded apparently higher survival rate as compared with those of slow cooling and rapid thawing and rapid cooling and slow thawing. Rapid cooling (at a rate of 0.3 deg. per minute to minus 40 deg. followed by immediate firect plunging into liquid nitrogen ) was also effective when combined with rapid thawing, or direct plunging into pre-warmed water bath at 37 deg. C. Murine oocytes aft
… More
e thawing were eveluated morphologically both with light and electron microscopes to reveal marked ultrastructural changes in vital organellas such as mitochondria being bulged and degenrated, in the oocytes which appered to be almost normal under light mictoscopic examination. As for human cases, five embryos at vatrious developmental stages and one unfertilized oocyte were frozen in 1.5 M dimethyl sulfoxide (DMSO) added in three steps at a cooling rate of 0.3 deg.C. per minute after nanual seeding procedure to nimus 40 deg.C. and then plunged directly into liquid nitrogen for storage. Following thwaing by dipping then into pre-warmed water at 37 deg.C., the cryoprotectant was temoved in five steps and cultured overnight. Morphological examination with light microscope reveald three embryos (two eight-cell embryos and one three-cell embryo) and an unfertilized oocyte, i.d., about 67 per cent of the cells were morphologically intact. Subsequent embryonsl transfer was not carried out according to an official regulation issued by The Japan Association of Obstetricvs and Gynecology. Less
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