Attempt at creating donor pig by nuclear transplantation for xenotrasplantation

异种移植用核移植培育供体猪的尝试

基本信息

批准号：
09671389
负责人：
TANIGUCHI Shigeki
金额：
$ 2.05万
依托单位：
Nara Medical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09671389/
关键词：
heart taransplantation xenotrasplantion gene transfer gene gun a 1-3 galactosiltransferase knockout EB-Virus episomal vector cryopreserved xenograft transplantation 凍結保存異種組織移植 α 1-3 galactosyltransferase knock out

项目摘要

1) My colleges and I have attempted to create alpha-gal knockout pig. Now, we are not successful in it yet. 2) To examine the means to change the allo- or xeno-reactivity after transplantation, we pefformed ex vivo transfer of marker gene to heart grafts using adenovirus vector. Successful gene transfer and expression of beta-gal gene were demonstrated. This result demonstrated that cardiomyocytes of the grafts expressed an exogenous gene products by adenovirus mediated gene transfer under hypothermic preservation condition. 3) We found that gene gun- mediated transfer of the EBV-based episomal vector into rat hearts results in long-lasting expression of a marker gene. Combined use of the gene gun and EBV-based episomal vector may contribute to gene therapy of cardiovascular diseases. To our knowledge, this is the first study showing that in vivo gene transfer into heart can be achieved using a gene gun. 4) We transfected rodent cells with EBV-episomal vectors carrying E.coli b-galactosidase gene as a marker gene. Significantly higher transient expression was demonstrated, both in vivo and in vitro, in rodent cells transfected with the EBV vector compared to the cells transfected with a conventional vector. 5) We transplanted porcine cryopreserved aortic valve to dog aorta for the purpose of assessment of the ability of discordant xeno-tissue-transplantation. Compared with fresh tissue, cryopreserved valves kept cell viability. This result suggests the ability of clinical use.

1）我和我的大学试图创建Alpha-Gal淘汰猪。现在，我们还没有成功。 2）要检查移植后改变同种或异种反应性的手段，我们使用腺病毒载体将标记基因转移到心脏移植物中。证明了β-GAL基因的成功基因转移和表达。该结果表明，在低温保存条件下，移植物的心肌细胞通过腺病毒介导的基因转移表达了外源基因产物。 3）我们发现基因枪介导的基于EBV的偶发载体转移到大鼠心脏中会导致标记基因的持久表达。基因枪和基于EBV的偶发载体的联合使用可能有助于心血管疾病的基因治疗。据我们所知，这是第一个研究表明，可以使用基因枪来实现体内基因转移到心脏。 4）我们用带有大肠杆菌b-半乳糖苷酶基因的EBV- eBISOMAL载体作为标记基因转染了啮齿动物细胞。与用常规载体转染的细胞相比，用体内和体外的瞬态表达明显更高。 5）我们将猪冷冻保存的主动脉瓣移植到狗主动脉中，以评估不一致的Xeno-Tissue转移的能力。与新鲜组织相比，冷冻保存的瓣膜保持细胞活力。该结果表明临床使用的能力。