The production of transgenic pigs for xenotransplantation

用于异种移植的转基因猪的生产

• 批准号: 10556059
• 负责人: SATO Eimei
• 金额: $ 8.06万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-10556059/
• 关键词: ES cells; nuclear transfer; embryo-derived cells; GFP; transgenic pigs; xenotransplantation; gene-knockout; somatic cell-clone; 遺伝子改革ブタ; 胚性幹細胞; 卵子; GFP発現細胞; 胚移植; ブタ; 異種移植; 遺伝子ノックアウトブタ; 発生工学

1) Basic technoloeies for the production of transgenic pigs, such as in vitro maturation and fertilization and culture of oocytes, nuclear transfer, freezing of embryos, embryo transfer, have been established.2) mitosen-activated protein kinase (MAPK) mediates the maturation-inducing signal from the cytoplasm into nucleus and induces meiosis reinitiation in porcine oocytes.3) The effect of hyaluronic acid on the development of in vitro-produced porcine embryos varies with the condition of oocyte maturation and fertilization.4) A porcine cell line can be established from the hatched blastocyst and maintained in vitro for a long period, and that reconstructed embryos obtained by transferring the blastocyst-derived cells into enucleated oocytes have the ability to develop to the blastocyst stage in vitro.5) Vector including a gene encoding green fluorescent protein (GFP gene) was developed, and techniques for selection of the cells with GFP gene was established .6) Techniques of nuclear transfer of the cells with GFP gene into enucleated oocytes was established.

1)转基因猪生产的基本技术，如卵母细胞的体外成熟、受精和培养、核移植、胚胎冷冻、胚胎移植、2）丝裂素活化蛋白激酶（mitosen-activated protein kinase，MAPK）介导成熟诱导信号从细胞质进入细胞核，诱导猪卵母细胞减数分裂重新启动; 3）透明质酸对猪卵母细胞体外发育的影响产生的猪胚胎随卵母细胞成熟和受精的条件而变化。4）从孵化的囊胚可以建立猪细胞系并在体外长期维持，并且通过将胚泡来源的细胞转移到去核卵母细胞中获得的重构胚胎具有体外发育到胚泡阶段的能力。5）包含编码绿色荧光蛋白的基因的载体6）建立了携带绿色荧光蛋白（GFP）基因的细胞向去核卵母细胞核移植的技术。

项目成果

佐藤英明: "ティッシュエンジニアリング組織工学の基礎と応用"名古屋大学出版会. 289 (1999)

佐藤英明：《组织工程基础与组织工程应用》名古屋大学出版社289（1999）。

佐々田 比呂志: "インテグリンと生殖内分泌"ホルモンと臨床. 47(7). 639-646 (1999)

Hiroshi Sasada：“整合素和生殖内分泌”激素和临床科学 47(7) (1999)。

佐藤英明: "Establishment of a porcine cell line from in vitro-produced blastocysts and transfer of the cells into enucleated oocytes"Biol.Reprod. 62. 1640-1646 (2000)

Hideaki Sato：“从体外产生的囊胚建立猪细胞系并将细胞转移到去核卵母细胞中”Biol.Reprod. 62. 1640-1646 (2000)

佐藤英明: "移植臓器生産トランスジェニックブタの開発-作出戦略と研究の現状"研究ジャーナル. 23. 15-20 (2000)

佐藤秀明：“用于移植器官生产的转基因猪的开发 - 生产策略和研究的现状”研究杂志。23. 15-20 (2000)。

Eimei Sato: "Development of a gene-modified pig using a cloning technology"Chemistry and Biology. 38. 508-513 (2000)

佐藤英明：“利用克隆技术开发基因改造猪”化学与生物学。