A chemically-defined and xeno-free culture system for derivation and in vivo chimeric cells tracking of monkey stem cell. Macaca fascicularis

Non-human primates are extremely valuable basic biology research tools, and are increasingly used in preclinical trials to test the safety and efficacy of biotechnology therapies. To reduce the need for this precious resource, it would be highly advantageous to use NHP cellular models in clinical studies. However, the generation of primate pluripotent stem cells (PSCs) is an undeveloped field, and the difficulty in detecting induced PSCs (iPSCs) limits the application of biotechnology in the cynomolgus monkey. Here, we report a chemically defined and xeno-free culture system for culturing and deriving monkey PSCs in vitro. The cells express hallmark naive pluripotency factors and additionally display features of global gene expression, genome-wide hypomethylation distinct from primed cells and found signaling pathways related with chimeric embryos that were potentially. Crucially for applications in biomedicine, we were also able to trace chimeric embryos in vivo and in vitro by integrating bioluminescent reporter genes into the monkey stem cells, as well as preserve their embryonic and extraembryonic developmental potentials. Meanwhile, we were generating a chimeric monkey with bioluminescence, which trace chimeric cells displayed more than one year for in monitoring in living animals. Therefore, our study could have broad utility in future applications of organoids from primate stem cells and chimeric monkey with bioluminescence in clinical studies.