Non-human primates (NHP) are increasingly used in preclinical trials to test the safety and efficacy of biotechnology therapies. Nonetheless, given the ethical issues and costs associated with this model, it would be highly advantageous to use NHP cellular models in clinical studies. However, developing and maintaining the naïve state of primate pluripotent stem cells (PSCs) remains difficult as does in vivo detection of PSCs, thus limiting biotechnology application in the cynomolgus monkey. Here, we report a chemically defined, xeno-free culture system for culturing and deriving monkey PSCs in vitro. The cells display global gene expression and genome-wide hypomethylation patterns distinct from monkey primed cells. We also found expression of signalling pathways components that may increase the potential for chimera formation. Crucially for biomedical applications, we were also able to integrate bioluminescent reporter genes into monkey PSCs and track them in chimeric embryos in vivo and in vitro. The engineered cells retained embryonic and extraembryonic developmental potential. Meanwhile, we generated a chimeric monkey carrying bioluminescent cells, which were able to track chimeric cells for more than two years in living animals. Our study could have broad utility in primate stem cell engineering and in utilizing chimeric monkey models for clinical studies.

中文翻译：

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非人类灵长类动物体内嵌合细胞的长期追踪。

非人类灵长类动物 (NHP) 越来越多地用于临床前试验，以测试生物技术疗法的安全性和有效性。尽管如此，考虑到与该模型相关的伦理问题和成本，在临床研究中使用 NHP 细胞模型将非常有利。然而，开发和维持灵长类多能干细胞（PSC）的初始状态仍然很困难，体内检测PSC也很困难，因此限制了生物技术在食蟹猴中的应用。在这里，我们报告了一种化学成分明确的、无异源的培养系统，用于体外培养和衍生猴 PSC。这些细胞表现出与猴子引发的细胞不同的全局基因表达和全基因组低甲基化模式。我们还发现信号通路成分的表达可能会增加嵌合体形成的可能性。对于生物医学应用至关重要的是，我们还能够将生物发光报告基因整合到猴子 PSC 中，并在体内和体外的嵌合胚胎中追踪它们。工程细胞保留了胚胎和胚胎外发育潜力。与此同时，我们生成了一只携带生物发光细胞的嵌合猴，它能够在活体动物中追踪嵌合细胞两年多。我们的研究在灵长类干细胞工程和利用嵌合猴模型进行临床研究方面具有广泛的用途。