Practical use of cloning technology for NOD and dy disease model mice production

克隆技术在NOD和DY疾病模型小鼠生产中的实际应用

• 批准号: 13558099
• 负责人: KONO Tomohiro
• 金额: $ 8.9万
• 依托单位: TOKYO UNIVERSITY OF AGRICULTURE
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13558099/
• 关键词: NOD mice; dy mice; mdx mice; embryo cloning; ES cells; disease model mice

The present study was conducted to develop embryo cloning systems using ES cell and contribute to fundamental studies for disease and its treatments. Here, we focused on NOD non-obese diabetic mouse used as a model for diabetes typeI, and dy and mdx mice used as muscular dystrophy and tried to produce cloned mice from ES cells derived from above disease model mice. The results obtained were summarized as follows. 1)By standard nuclear transfer, cloned blastocysts were successfully produced using ES cells established from NOD mice, but no pup was obtained to date. 2)ES cells, which can be cultured over several passages, have been established from dy and mdx mice. 3)Using these ES cells as donor nuclei, cloned blastocysts were produced, but again no pup was obtained. Together, we have established ES cells from NOD, dy and mdx mice, but failed to produce cloned individuals. Further studies may be necessary to clarify developmental competence of ES cells established here.

本研究旨在利用胚胎干细胞建立胚胎克隆系统，为疾病及其治疗的基础研究做出贡献。本研究以NOD非肥胖糖尿病小鼠作为糖尿病ⅰ型模型，dy和mdx小鼠作为肌肉萎缩症模型，并尝试用上述疾病模型小鼠衍生的ES细胞制备克隆小鼠。所得结果总结如下：1)通过标准核移植，利用NOD小鼠胚胎干细胞成功制备了克隆囊胚，但尚未获得幼仔。2)从dy和mdx小鼠身上建立了可多次培养的胚胎干细胞。3)利用这些胚胎干细胞作为供核，获得了克隆囊胚，但仍未获得幼仔。我们一起从NOD、dy和mdx小鼠身上建立了胚胎干细胞，但未能产生克隆个体。可能需要进一步的研究来阐明这里建立的胚胎干细胞的发育能力。

项目成果

河野友宏: "ES細胞を用いたクローンマウス"医学のあゆみ. 203. 1061-1064 (2002)

Tomohiro Kono：“使用 ES 细胞克隆小鼠”医学史 203. 1061-1064 (2002)。

Nagafuchi S.: "Epstein-Barr virus survival : expression and release of Fas ligand."Intern.Med.. 41. 603-604 (2002)

Nagafuchi S.：“Epstein-Barr 病毒存活：Fas 配体的表达和释放。”Intern.Med.. 41. 603-604 (2002)

Sotomaru Y, Katsuzawa Y, Domeki I, Kono T.: "Determination by real-time RT-PCR of imprinted expression of the insulin-like growth factor II(Igf2)gene in mouse uniparental fetuses."J.Reprod.Dev.. 47. 139-144 (2001)

Sotomaru Y、Katsuzawa Y、Domeki I、Kono T.：“通过实时 RT-PCR 测定小鼠单亲胎儿中胰岛素样生长因子 II (Igf2) 基因的印迹表达。”J.Reprod.Dev..

Ogawa H, Ono Y, Shimozawa N, Sotomaru Y, Hiura H, Ito M, Kono T.: "Disruption of imprinting in cloned mouse fetuses from embryonic stem cells"Reproduction. 126. 549-557 (2003)

Okawa H、Ono Y、Shimozawa N、Sotomaru Y、Hiura H、Ito M、Kono T.：“胚胎干细胞克隆小鼠胎儿印记的破坏”复制。

Shimozawa, N., Ono, Y., Muguruma, K., Hioki, K., Arai, Y., Kono, T., Ito, M.: "Direct production of gene-targeted mice from ES cells by nuclear transfer and gene transmission to their progeny"Exp. Anim.. 51. 375-381 (2002)

Shimozawa, N.、Ono, Y.、Muguruma, K.、Hioki, K.、Arai, Y.、Kono, T.、Ito, M.：“通过核移植和胚胎干细胞直接生产基因靶向小鼠