Genetic manipulation and transplantation of cynomolgus embryonic stem cells

将遗传物质稳定地引入灵长类胚胎干细胞（ES）的能力可能会有更广泛的应用。在这项研究中，我们研究了慢病毒基因在食蟹ES细胞中的转移。用编码绿色荧光蛋白（GFP）基因的基于猴免疫缺陷病毒（SIV）的慢病毒载体转染食蟹蟹ES细胞一次后，大多数细胞（约90%）发出荧光，并且高水平的GFP表达持续了5个月而无需选择程序。此外，在胚状体形成过程中观察到高水平的GFP表达。另一方面，基于SIV的载体转导小鼠胚胎干细胞的基因转导率较低，这表明灵长类病毒载体转导灵长类胚胎干细胞的效率高于小鼠胚胎干细胞。使用GFP作为报告基因，可以直接和简单地检测成功转导的胚胎干细胞，并有助于在体外和体内监测胚胎干细胞的增殖和分化。此外，这种高效的基因转移方法允许忠实地将基因传递到灵长类胚胎干细胞，具有研究和治疗应用的潜力。

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.