Electroporation of cynomolgus monkey embryonic stem cells
Electroporation of cynomolgus monkey embryonic stem cells
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DOI：
10.1002/gene.10246
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发表时间：
2003-12-01
期刊：
影响因子：
1.5
通讯作者：
Suemori, H
中科院分区：
文献类型：
--
作者：
Furuya, M;Yasuchika, K;Suemori, H
灵长类动物胚胎干细胞的高效遗传修饰是基础研究和临床前研究的基础。据报道，灵长类动物ES细胞的转染效率低于小鼠ES细胞。食蟹猴ES细胞为了解人类发育和疾病提供了一个强大的模型。我们评估了电穿孔作为将外源基因导入食蟹猴ES细胞的方法。我们的检查允许我们建立从10（7）个食蟹猴ES细胞产生约100个稳定转染的克隆的方案。然而，在其他ES细胞系中观察到效率的差异。我们比较了PGK-1，CMV和SV 40启动子在产生有效G418选择的食蟹猴ES细胞中的转录活性。虽然PGK-1和SV 40启动子有效地驱动neo基因表达，但CMV启动子的转录活性明显较低。在食蟹猴ES细胞中。利用这种电穿孔方法，我们建立了荧光食蟹猴ES细胞系。这些细胞可能是有用的工具，用于跟踪移植的细胞在移植研究中使用的各种功能细胞来源于食蟹猴ES细胞。(C)2003 Wiley-Liss，Inc.
Efficient genetic modification of primate embryonic stem (ES) cells is essential for the application for both basic and preclinical research. The transfection efficiency of primate ES cells is reportedly lower than that of mouse ES cells. Cynomolgus monkey ES cells provide a powerful model for understanding human development and disease. We evaluated electroporation as a method to introduce foreign genes into cynomolgus monkey ES cells. Our examination has allowed us to establish a protocol producing about 100 stably transfected clones from 10(7) cynomolgus monkey ES cells. Differences in efficiency, however, were observed for other ES cell lines. We compared the transcriptional activities of the PGK-1, CMV, and SV40 promoters in cynomolgus monkey ES cells generating efficient G418 selection. Although the PGK-1 and SV40 promoters efficiently drove neo gene expression, the CMV promoter was significantly less transcriptionally active. in cynomolgus monkey ES cells. Using this electroporation method, we established fluorescent cynomolgus monkey ES cell lines. These cells may be useful tools for tracing grafted cells in transplantation studies using a variety of functional cells derived from cynomolIgus monkey ES cells. (C) 2003 Wiley-Liss, Inc.