DEVELOPMENT OF THE SYSTEM FOR CREATING A TRANSGENIC MINIPIG WITH THE THREE-DIMENSIONAL MONITORING SYSTEM

三维监测系统转基因小型猪创建系统的开发

• 批准号: 08508003
• 负责人: TSUJI Takayuki
• 金额: $ 27.01万
• 依托单位: NATIONAL CARDIOVASCULAR CENTER RESEARCH INSTITUTE
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08508003/
• 关键词: transgenic minipig; three-dimensional rigid endoscope; ova-holding device; computer-guided microinjector; microporous glass plate; gene engineering

Aim of the project is to establish an efficient system of creating transgenic minipigs for heterogenic organ transplantation. We invented three-dimensional (3D) rigid endoscopes for obtaining less-invasively minipig ova from ovarium without laparotomy and an ova-holding device having 5 x 5 micropocket matrix made of microporous glass plate. The device can suck ova into micropockets having diameter of 150 micrometer with solution circulating across the plate fixed in the holder housing. Ova can be cultured in the device under controlled environments of appriate temperature, pH, and gas contents. GFP marked-human gene can be introduced into the nucleus with the computer guided microinjector. All the procedures can be monitored with the 3 D microscope and recorded with a video-cassette recorder. The cultured GFP- marked embryo can be selectively picked up and introduced laparoscopically into female minipig uterus less-invasively with the 3 D rigid endoscope. This system can create several ten times more efficiently a trangenic minipig than the conventional method which needs much skill and more female minipig sacrification.

该项目的目的是建立一个有效的系统来创建用于异源器官移植的转基因小型猪。我们发明了三维 (3D) 刚性内窥镜，用于在不进行剖腹手术的情况下从卵巢中以微创方式获取小型猪卵子，以及具有由微孔玻璃板制成的 5 x 5 微囊矩阵的卵子保持装置。该装置可以将卵子吸入直径为 150 微米的微囊中，溶液在固定在支架外壳中的板上循环。卵子可以在适当温度、pH 和气体含量的受控环境下在设备中培养。可以用计算机引导的显微注射器将GFP标记的人类基因引入细胞核。所有的过程都可以用 3D 显微镜监控并用录像机记录。培养的 GFP 标记胚胎可以通过 3D 刚性内窥镜选择性拾取并通过腹腔镜微创导入雌性小型猪子宫。该系统可以比传统方法高效数十倍地生产转基因小型猪，传统方法需要更多的技巧和更多的雌性小型猪的牺牲。

项目成果

Tsuji T, et al: "Microinjection monitoring of minipig fertilized eggs with 3-D microscopes" Proceedings of lntl Conf on New Frontiers in Biomechanical Engineering. 381-382 (1997)

Tsuji T 等人：“使用 3D 显微镜显微注射监测小型猪受精卵”生物力学工程新前沿国际会议论文集。

井内康浩,辻 隆之,他: "マイクロポーラスセラミックスを応用した流れ場" 第7回バイオエンジニアリング学術講演会講演論文集. 96-29. 158-159 (1996)

Yasuhiro Inouchi、Takayuki Tsuji 等：“使用微孔陶瓷的流场”第七届生物工程学术会议论文集 96-29（1996）。

Tsuji T, Fujisato T, Miyawaki F, e al.: "Microinjection monitoring of minipig fertilized eggswith 3-D microscopes" Proceedings of Intl Conf on New Frontiers in Biomechanical Engineering. 381-382 (1997)

Tsuji T、Fujisato T、Miyawaki F 等人：“用 3D 显微镜显微注射监测小型猪受精卵”生物力学工程新前沿国际会议论文集。

Fujisato T,Iuchi Y,et al: "Development of fixation device of fertilized ova for gene injection" Proceedings of Intl Conf on New Frontiers in Biomech Engineering. 379-380 (1997)

Fujisato T、Iuchi Y 等人：“用于基因注射的受精卵固定装置的开发”生物力学工程新前沿国际会议论文集。

Tsuji T,et.al: "Microinjection monitoring of minipig fertilized eggs with 3-D microscopes" Proceedings of Intl Conf on New Frontiers in Biomechanical Engineering. 381-382 (1997)

Tsuji T 等人：“使用 3D 显微镜显微注射监测小型猪受精卵”生物力学工程新前沿国际会议论文集。