Abstract
Non-human primates are our closest relatives and are of special interest for ecological, evolutionary and biomedical research. The Japanese macaque (Macaca fuscata) has contributed to the progress of primatology and neurosciences over 60 years. Despite this importance, the molecular and cellular basis of the Japanese macaque remains unexplored since useful cellular tools are lacking. Here we generated induced pluripotent stem cells (iPSCs) from skin fibroblasts of the Japanese macaque with Sendai virus or plasmid vectors. The Japanese macaque iPSCs (jm-iPSCs) were established under feeder-free culture conditions, but feeder cells turned out to be essential for their maintenance. The jm-iPSCs formed human iPSC-like flat colonies which were positive for pluripotent antigens including alkaline phosphatase, SSEA4, and TRA-1-81. They also expressed endogenous OCT3/4, SOX2, L-MYC, and KLF4 and other pluripotent marker genes. The potential to differentiate into all three germ layers and neural stem cells was confirmed by embryoid body and neurosphere formation, respectively. The jm-iPSCs will provide a robust in vitro tool for investigating the underlying mechanisms of development and physiology studies with the Japanese macaque.
Highlights
The Japanese macaque (Macaca fuscata) is one species of the Asian macaque monkeys along with rhesus and cynomolgus monkeys
The jm-induced pluripotent stem cells (iPSCs) were similar with human iPSCs in colony morphology, marker gene expression, and differentiation potency albeit their growth totally depended on feeder cells
Human POU5F1, SOX2, KLF4, and L-MYC were transduced into jm-fibroblasts by Sendai virus (SeV) infection, while plasmid vectors bearing human OCT3/4, SOX2, KLF4, LIN28, L-MYC, and shRNA for TP53 were transfected with an additional EBNA1 plasmid by lipofection
Summary
The Japanese macaque (Macaca fuscata) is one species of the Asian macaque monkeys along with rhesus and cynomolgus monkeys. The Japanese macaque has been employed as an excellent primate model with the accumulated knowledge and a potential to integrate the laboratory studies with the field research. Difficulties in developmental physiology and genetic manipulations in vivo restrict in-depth analyses of molecular and cellular mechanisms in the Japanese macaque biology. Induced pluripotent stem cells (iPSCs) can provide a new strategy to compensate this experimental concern of the Japanese macaque. IPSCs have a potential to differentiate into all three germ layers (ectoderm, mesoderm, and endoderm) and germ cells both in vivo and in vitro. IPSCs can serve as a potent in vitro tool to address the molecular and cellular basis of development and physiology in the Japanese macaque. The jm-iPSCs will be a powerful in vitro counterpart to facilitate the Japanese macaque biology
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