Roodgar, Morteza, Suchy, Fabian P., Nguyen, Lan H., Bajpai, Vivek K., Sinha, Rahul, Vilches-Moure, Jose G., Van Bortle, Kevin, Bhadury, Joydeep, Metwally, Ahmed, Jiang, Lihua, Jian, Ruiqi, Chiang, Rosaria, Oikonomopoulos, Angelos, Wu, Joseph C., Weissman, Irving L., Mankowski, Joseph L., Holmes, Susan, Loh, Kyle M., Nakauchi, Hiromitsu, and VandeVoort, Catherine A.
As our closest living relatives, non-human primates uniquely enable explorations of human health, disease, development, and evolution. Considerable effort has thus been devoted to generating induced pluripotent stem cells (iPSCs) from multiple non-human primate species. Here, we establish improved culture methods for chimpanzee (Pan troglodytes) and pig-tailed macaque (Macaca nemestrina) iPSCs. Such iPSCs spontaneously differentiate in conventional culture conditions, but can be readily propagated by inhibiting endogenous WNT signaling. As a unique functional test of these iPSCs, we injected them into the pre-implantation embryos of another non-human species, rhesus macaques (Macaca mulatta). Ectopic expression of gene BCL2 enhances the survival and proliferation of chimpanzee and pig-tailed macaque iPSCs within the pre-implantation embryo, although the identity and long-term contribution of the transplanted cells warrants further investigation. In summary, we disclose transcriptomic and proteomic data, cell lines, and cell culture resources that may be broadly enabling for non-human primate iPSCs research. [Display omitted] • Inhibiting WNT signaling improves culture of chimp and pig-tailed macaque iPSCs • Endogenous high expression of BCL2 in the rhesus inner cell mass • Expression of BCL2 enhances proliferation of chimp iPSCs in rhesus early embryo • Co-culture of chimp and human iPSCs can differentiate into integrated cardiomyocytes Roodgar et al. demonstrate an improved culture of chimpanzee and pig-tailed macaque iPSCs. They show high expression of endogenous BCL2 in rhesus macaque ICM. Through over expression of BCL2 in chimpanzee iPSCs, they generate primate cross-species pre-implantation embryos composed of chimpanzee iPSCs and rhesus pre-implantation embryos. [ABSTRACT FROM AUTHOR]