@article{9ceaccb1083443ef9eaabf952eec4e78,
title = "Chimpanzee and pig-tailed macaque iPSCs: Improved culture and generation of primate cross-species embryos",
abstract = "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.",
keywords = "CP: Developmental biology, Nonhuman primates, blastocyst, BCL2, chimpanzee, cross-species embryos, iPSCs, interspecies chimera, pig-tailed macaque",
author = "Morteza Roodgar and Suchy, {Fabian P.} and Nguyen, {Lan H.} and Bajpai, {Vivek K.} and Rahul Sinha and Vilches-Moure, {Jose G.} and {Van Bortle}, Kevin and Joydeep Bhadury and Ahmed Metwally and Lihua Jiang and Ruiqi Jian and Rosaria Chiang and Angelos Oikonomopoulos and Wu, {Joseph C.} and Weissman, {Irving L.} and Mankowski, {Joseph L.} and Susan Holmes and Loh, {Kyle M.} and Hiromitsu Nakauchi and VandeVoort, {Catherine A.} and Snyder, {Michael P.}",
note = "Funding Information: Support comes from a gift to the Stanford Department of Genetics and the Stanford Center for Genomics and Personalized Medicine (SCGPM), the California National Primate Research Center (CNPRC) grant P51OD011107, the Johns Hopkins University (JHU) grant 5U42OD013117 for the pigtailed macaque colony, and the Southwest National Primate Research Center grant P51OD011133 from the office of Research Infrastructure Programs, National Institutes of Health. We would like to thank David Glenn Smith for his continuing support in consulting on NHP species selection and primate evolutionary biology that apply to the study design and PSC biology. We would like to thank Drs. John Morrison and Jeff Roberts at the CNPRC for their support and for facilitating the animal experiments. We also thank Koosha Nassiri Nazif and Mahsa Roodgar for assisting in additional image analysis, including the use of KLayout and MATLAB, and Tuhin K. Guha for assisting in imaging mouse tissue slides. M.R. is supported by the National Institutes of Health, National Heart, Lung, and Blood Institute (NHLBI), Stanford training grant T32HL120824-6. K.V.B. is supported by the National Institutes of Health, National Human Genome Research Institute (NHGRI) grant K99HG010362. V.K.B. is supported by grant no. CA160997. K.M.L. is a Packard Foundation Fellow, Pew Scholar, Human Frontier Science Program Young Investigator, Baxter Foundation Faculty Scholar, and The Anthony DiGenova Endowed Faculty Scholar. M.R. conceived the study, species selection, iPSC derivation, and NHP iPSC media optimization. K.M.L. guided us on iPSC medium optimization. M.R. and F.P.S. conducted the iPSC fluorescent labeling, single-cell expansion, and BCL2 transduction. C.A.V. conducted rhesus macaque oocyte retrieval, in vitro fertilization (IVF), and embryo generation. F.P.S. conducted the embryo injection and imaging. M.R. F.P.S. and J.B. conducted the iPSC co-culture experiments. M.R. V.K.B. and A.O. conducted the cardiomyocyte differentiation and evaluation. M.R. L.H.N. R.C. K.V.B. and R.S. conducted the transcriptome experiments and analysis. L.J. R.J. A.M. and M.R. conducted the proteomics experiments and analysis. M.R. L.H.N. A.M. and R.S. conducted the transcriptome and proteomics analysis. V.K.B. conducted the ectoderm differentiation. M.R. and J.G.V.-M. conducted the BCL2 teratoma assays and histopathology analysis. J.L.M. supervised the species selection. K.V.B. and J.L.M. edited the manuscript. J.C.W. supervised the cardiomyocyte experiments. C.A.V. H.N. and M.P.S. supervised the study. The authors declare no competing interests. Funding Information: Support comes from a gift to the Stanford Department of Genetics and the Stanford Center for Genomics and Personalized Medicine (SCGPM), the California National Primate Research Center ( CNPRC ) grant P51OD011107 , the Johns Hopkins University ( JHU ) grant 5U42OD013117 for the pigtailed macaque colony, and the Southwest National Primate Research Center grant P51OD011133 from the office of Research Infrastructure Programs, National Institutes of Health . We would like to thank David Glenn Smith for his continuing support in consulting on NHP species selection and primate evolutionary biology that apply to the study design and PSC biology. We would like to thank Drs. John Morrison and Jeff Roberts at the CNPRC for their support and for facilitating the animal experiments. We also thank Koosha Nassiri Nazif and Mahsa Roodgar for assisting in additional image analysis, including the use of KLayout and MATLAB, and Tuhin K. Guha for assisting in imaging mouse tissue slides. M.R. is supported by the National Institutes of Health , National Heart, Lung, and Blood Institute ( NHLBI ), Stanford training grant T32HL120824-6 . K.V.B. is supported by the National Institutes of Health , National Human Genome Research Institute ( NHGRI ) grant K99HG010362 . V.K.B. is supported by grant no. CA160997 . K.M.L. is a Packard Foundation Fellow, Pew Scholar, Human Frontier Science Program Young Investigator, Baxter Foundation Faculty Scholar, and The Anthony DiGenova Endowed Faculty Scholar. Publisher Copyright: {\textcopyright} 2022 The Authors",
year = "2022",
month = aug,
day = "30",
doi = "10.1016/j.celrep.2022.111264",
language = "English (US)",
volume = "40",
journal = "Cell Reports",
issn = "2211-1247",
publisher = "Cell Press",
number = "9",
}