1992 Fiscal Year Annual Research Report

哺乳動物における受精卵(胚)の体外操作(凍結、分離、融合)に関する研究

Research Project

Project/Area Number	02556039
Research Institution	Hokkaido University
Principal Investigator	金川弘司北海道大学, 獣医学部, 教授 (00111162)
Co-Investigator(Kenkyū-buntansha)	石森久雄雪印乳業(株)受精卵移植研究所, 研究員清家昇雪印乳業(株)受精卵移植研究所, 主席研究員菱沼貢北海道大学, 獣医学部, 助手 (30183578) 高橋芳幸北海道大学, 獣医学部, 助教授 (70167485)
Keywords	受精卵(胚) / 凍結保存 / ガラス化 / 胚の融合 / 核移植
Research Abstract	胚の凍結に関する研究では、エチレングリコール(EG)と糖を用いた急速凍結凍結に関して、3mMのEGと0.25mMのトレハロースあるあはスクロースの混合溶液はマウス未受精卵子の凍結に有効であり、約60%の生存率が得られ、ガラス化卵子の体外受精率、その後の発育ならびに移植後の産子への発育成績は未凍結卵子の成績に等しいことが判った。ガラス化に関しては、25%エチレングリコールと25%DMSOの混合溶液あるいは20%エチレングリコール、20%DMSOおよび10%1-3ブタンダイオールの混合溶液は、マウス桑実胚ならびに胚盤胞では90%以上の生存率が得られ、さらに、これら2種類のガラス化溶液は、ウシ桑実胚および胚盤胞のガラス化に有効であり、ガラス化したウシ胚を移植して多数の産子が得られることを実証した。胚の分離および融合技術を応用した胚のクローニングに関する研究では、マウス胚を用いて各種の基礎的な検討を行い、多くの知見を得た。とくに、除核した未受精卵子に核体を移植して作出した核移植胚の初期化にとって、核体の細胞周期が重要であることを見い出した。すなわち、細胞周期の初期ステージにある細胞核を移植することにより、初期化が高率に起こることが判明し、初期の細胞周期にある4および8細胞期胚の細胞核を移植した核移植胚から世界で初めてのクローンマウスの作出に成功した。

Research Products
(14 results)

All Other

All Publications (14 results)

[Publications] Rayos AA: "Quick freezing of mouse two-,four-,and eight-cell embryos with ethylene glycol plus sucrose or lactose:effects of developmental stage and equilibration period on survival in vitro" Anim Reprod Sci. 27. 239-245 (1992)
[Publications] Cheong HT: "Influences of electrode geometry and field strength on the fusion and subsequent development of mouse two-cell embryos." Anim Reprod Sci. 29. 307-317 (1992)
[Publications] Cheong HT: "Development of mouse embryonic nulei transferred to enucleated oocytes and zygotes." Jpn.J.Vet.Res. 40. 149-159 (1992)
[Publications] Cheon HT: "Developmental capacity of reconstituted mouse embryos:influences of nuceus and cytoplasm souces." J.Vet.Med.Sci. 54. 1099-1103 (1992)
[Publications] Ishimori H: "Viability of vitrified mouse embryos using various cryoprotectant mixtures." Theiogenology. 37. 481-487 (1992)
[Publications] Rayos AA: "Quick freezing of one-cell mouse embryos using ethylene glycol with sucrose." Theriogenology. 37. 595-603 (1992)
[Publications] Taniguchi T: "Development of reconstituted mouse embryos produced from bisected electrofused pronuclear-stage embryos." Theriogenology. 37. 655-663 (1992)
[Publications] Taniguchi T: "Development of reconstituted mouse embryos produced from the cytoplast of bisected oocytes or pronuclear-stage embryosand single balstomeres of 2-cell stage embryos." Theriogenology. 38. 921-934 (1992)
[Publications] Ishimori H: "Factors affecting survial of mouse blastocysts vitrified by a mixture of ethylene glycol and demithy sulfoxide." Theriogenology. 38. 1175-1185 (1992)
[Publications] Valdez CA: "Crypreservation of mouse embryos using a plant vitrification solution." Proc 8th Conger Fed Asian Vet.Assoc. 368-372 (1992)
[Publications] Kanagawa.H: "Embryo cryopreservation using simplified freezing techniques." Proc.8th Conger.Fed.Asian Vet.Assoc. 547-553 (1992)
[Publications] 藤川清三: "マウス胚の一段ガラス化保存のメカニズム" 凍結及び乾燥研究会会誌. 38. 33-37 (1992)
[Publications] Valdez CA: "Successful cryopreservation of mouse blastocysts using a new vitrification solution." J.Reprod.Fertil. 96. 793-802 (1992)
[Publications] Cheong HT: "Birth of mice after transplanatation of early cell-cyclestage embryonic nuclei into oocytes." Biol Reprod. 48. (1993)

1992 Fiscal Year Annual Research Report

哺乳動物における受精卵(胚)の体外操作(凍結、分離、融合)に関する研究

Principal Investigator

金川 弘司 北海道大学, 獣医学部, 教授 (00111162)

Research Products

[Publications] Rayos AA: "Quick freezing of mouse two-,four-,and eight-cell embryos with ethylene glycol plus sucrose or lactose:effects of developmental stage and equilibration period on survival in vitro" Anim Reprod Sci. 27. 239-245 (1992)

[Publications] Cheong HT: "Influences of electrode geometry and field strength on the fusion and subsequent development of mouse two-cell embryos." Anim Reprod Sci. 29. 307-317 (1992)

[Publications] Cheong HT: "Development of mouse embryonic nulei transferred to enucleated oocytes and zygotes." Jpn.J.Vet.Res. 40. 149-159 (1992)

[Publications] Cheon HT: "Developmental capacity of reconstituted mouse embryos:influences of nuceus and cytoplasm souces." J.Vet.Med.Sci. 54. 1099-1103 (1992)

[Publications] Ishimori H: "Viability of vitrified mouse embryos using various cryoprotectant mixtures." Theiogenology. 37. 481-487 (1992)

[Publications] Rayos AA: "Quick freezing of one-cell mouse embryos using ethylene glycol with sucrose." Theriogenology. 37. 595-603 (1992)

[Publications] Taniguchi T: "Development of reconstituted mouse embryos produced from bisected electrofused pronuclear-stage embryos." Theriogenology. 37. 655-663 (1992)

[Publications] Taniguchi T: "Development of reconstituted mouse embryos produced from the cytoplast of bisected oocytes or pronuclear-stage embryosand single balstomeres of 2-cell stage embryos." Theriogenology. 38. 921-934 (1992)

[Publications] Ishimori H: "Factors affecting survial of mouse blastocysts vitrified by a mixture of ethylene glycol and demithy sulfoxide." Theriogenology. 38. 1175-1185 (1992)

[Publications] Valdez CA: "Crypreservation of mouse embryos using a plant vitrification solution." Proc 8th Conger Fed Asian Vet.Assoc. 368-372 (1992)

[Publications] Kanagawa.H: "Embryo cryopreservation using simplified freezing techniques." Proc.8th Conger.Fed.Asian Vet.Assoc. 547-553 (1992)

[Publications] 藤川 清三: "マウス胚の一段ガラス化保存のメカニズム" 凍結及び乾燥研究会会誌. 38. 33-37 (1992)

[Publications] Valdez CA: "Successful cryopreservation of mouse blastocysts using a new vitrification solution." J.Reprod.Fertil. 96. 793-802 (1992)

[Publications] Cheong HT: "Birth of mice after transplanatation of early cell-cyclestage embryonic nuclei into oocytes." Biol Reprod. 48. (1993)

金川弘司北海道大学, 獣医学部, 教授 (00111162)

[Publications] 藤川清三: "マウス胚の一段ガラス化保存のメカニズム" 凍結及び乾燥研究会会誌. 38. 33-37 (1992)