Genetic manipulation and transplantation of cynomolgus embryonic stem cells

食蟹猴胚胎干细胞的基因操作与移植

• 批准号: 13671080
• 负责人: HANAZONO Yutaka
• 金额: $ 2.62万
• 依托单位: Jichi Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671080/
• 关键词: embryonic stem cells; lentiviral vector; cynomolgus monkeys; primate model; NOD; SCID; 胎仔移植; カニクイザル; ヒツジ; GFP; OP9; BMP-4; ヒツジ胎仔

The ability to stably introduce genetic material into primate embryonic stem (ES) cells could allow broader application. In this study, we examined lentiviral gene transfer into cynomolgus ES cells. When cynomolgus ES cells were transduced once with a simian immunodeficiency virus (SIV)-based lentivirus vector encoding the green fluorescent protein (GFP) gene, most cells (around 90%) fluoresced and high levels of GFP expression persisted for 5 months without selection procedures. In addition, high levels of GFP expression were observed during embryoid body formation. On the other hand, transduction of mouse ES cells with the SIV based vector resulted in lower gene transfer rates, implying that primate viral vectors can transduce primate ES cells more efficiently than mouse ES cells. The use of GFP as a reporter gene allows direct and simple detection of successfully transduced ES cells and facilitates monitoring of ES cell proliferation and differentiation both in vitro and potentially in vivo. Furthermore, this highly efficient gene transfer method allows faithful gene delivery to primate ES cells with potential for both research and therapeutic application.

将遗传物质稳定引入灵长类胚胎干（ES）细胞的能力可能会带来更广泛的应用。在这项研究中，我们检查了慢病毒基因转移到食蟹猴 ES 细胞中的情况。当食蟹猴 ES 细胞用编码绿色荧光蛋白 (GFP) 基因的基于猿猴免疫缺陷病毒 (SIV) 的慢病毒载体转导一次时，大多数细胞（约 90%）发出荧光，并且高水平的 GFP 表达持续 5 个月，无需选择程序。此外，在拟胚体形成过程中观察到高水平的 GFP 表达。另一方面，用基于SIV的载体转导小鼠ES细胞导致较低的基因转移率，这意味着灵长类病毒载体可以比小鼠ES细胞更有效地转导灵长类ES细胞。使用 GFP 作为报告基因可以直接、简单地检测成功转导的 ES 细胞，并有助于监测体外和潜在体内 ES 细胞的增殖和分化。此外，这种高效的基因转移方法可以将基因忠实地传递到灵长类动物胚胎干细胞中，具有研究和治疗应用的潜力。

项目成果

Ageyama,N., et al.: "Safe and efficient methods of autologous hematopoietic stem cell transplantation for biomedical research in cynomolgus monkeys"Comp.Med.. 52. 445-451 (2002)

Ageyama,N. 等人：“用于食蟹猴生物医学研究的自体造血干细胞移植的安全有效的方法”Comp.Med.. 52. 445-451 (2002)

Hanazono,Y., et al.: "In vivo selective expansion of gene-modified hematopoietic cells in a nonhuman primate model"Gene Ther.. 9. 1055-1064 (2002)

Hanazono,Y., et al.：“非人灵长类动物模型中基因修饰造血细胞的体内选择性扩增”Gene Ther.. 9. 1055-1064 (2002)

Hanazono, Y.et al.: "Genetic manipulation of primate embryonic and hematopoietic stem cells with simian lentivirus vectors"Trends in Cardiovascular Medicine. 13巻. 106-110 (2003)

Hanazono, Y. 等人：“用猿猴慢病毒载体对灵长类胚胎和造血干细胞进行基因操作”，《心血管医学趋势》，第 13 卷，106-110（2003 年）。

Hanazono Y, et al.: "Genetic manipulation of primate embryonic and hematopoietic stem cells with simian lentivirus vectors"Trends in Cardiovascular Medicine. 13. 106-110 (2003)

Hanazono Y 等人：“用猿猴慢病毒载体对灵长类胚胎和造血干细胞进行基因操作”心血管医学趋势。

Wang, L. et al.: "Delayed delivery of AAV-GDNF prevents nigral neurodegeneration and promotes functional recovery in a rat model of Parkinson's disease"Gene Ther.. 9巻. 381-389 (2002)

Wang, L. 等人：“延迟递送 AAV-GDNF 可预防帕金森病大鼠模型中的黑质神经变性并促进功能恢复”Gene Ther.. 9. 381-389 (2002)