{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,31]],"date-time":"2026-03-31T05:32:19Z","timestamp":1774935139882,"version":"3.50.1"},"reference-count":46,"publisher":"Springer Science and Business Media LLC","issue":"8","license":[{"start":{"date-parts":[[2021,7,2]],"date-time":"2021-07-02T00:00:00Z","timestamp":1625184000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/www.springer.com\/tdm"},{"start":{"date-parts":[[2021,7,2]],"date-time":"2021-07-02T00:00:00Z","timestamp":1625184000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.springer.com\/tdm"}],"funder":[{"DOI":"10.13039\/100012636","name":"New York State Stem Cell Science","doi-asserted-by":"publisher","award":["C028129"],"award-info":[{"award-number":["C028129"]}],"id":[{"id":"10.13039\/100012636","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100012636","name":"New York State Stem Cell Science","doi-asserted-by":"publisher","award":["C029556"],"award-info":[{"award-number":["C029556"]}],"id":[{"id":"10.13039\/100012636","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100012636","name":"New York State Stem Cell Science","doi-asserted-by":"publisher","award":["C30290GG"],"award-info":[{"award-number":["C30290GG"]}],"id":[{"id":"10.13039\/100012636","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Nat Protoc"],"published-print":{"date-parts":[[2021,8]]},"DOI":"10.1038\/s41596-021-00565-7","type":"journal-article","created":{"date-parts":[[2021,7,2]],"date-time":"2021-07-02T16:05:56Z","timestamp":1625241956000},"page":"3954-3980","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Generation of mouse\u2013human chimeric embryos"],"prefix":"10.1038","volume":"16","author":[{"given":"Boyang","family":"Zhang","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hanqin","family":"Li","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhixing","family":"Hu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Houbo","family":"Jiang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Aimee B.","family":"Stablewski","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3547-1182","authenticated-orcid":false,"given":"Brandon J.","family":"Marzullo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Donald A.","family":"Yergeau","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7630-8800","authenticated-orcid":false,"given":"Jian","family":"Feng","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2021,7,2]]},"reference":[{"key":"565_CR1","doi-asserted-by":"publisher","first-page":"196","DOI":"10.1038\/nature05972","volume":"448","author":"PJ Tesar","year":"2007","unstructured":"Tesar, P. J. et al. New cell lines from mouse epiblast share defining features with human embryonic stem cells. Nature 448, 196\u2013199 (2007).","journal-title":"Nature"},{"key":"565_CR2","doi-asserted-by":"publisher","first-page":"1145","DOI":"10.1126\/science.282.5391.1145","volume":"282","author":"JA Thomson","year":"1998","unstructured":"Thomson, J. A. et al. Embryonic stem cell lines derived from human blastocysts. Science 282, 1145\u20131147 (1998).","journal-title":"Science"},{"key":"565_CR3","doi-asserted-by":"publisher","first-page":"487","DOI":"10.1016\/j.stem.2009.05.015","volume":"4","author":"J Nichols","year":"2009","unstructured":"Nichols, J. & Smith, A. Naive and primed pluripotent states. Cell Stem Cell 4, 487\u2013492 (2009).","journal-title":"Cell Stem Cell"},{"key":"565_CR4","doi-asserted-by":"publisher","first-page":"255","DOI":"10.1038\/309255a0","volume":"309","author":"A Bradley","year":"1984","unstructured":"Bradley, A., Evans, M., Kaufman, M. H. & Robertson, E. Formation of germ-line chimaeras from embryo-derived teratocarcinoma cell lines. Nature 309, 255\u2013256 (1984).","journal-title":"Nature"},{"key":"565_CR5","doi-asserted-by":"publisher","first-page":"90","DOI":"10.1016\/j.ydbio.2006.03.026","volume":"295","author":"D James","year":"2006","unstructured":"James, D., Noggle, S. A., Swigut, T. & Brivanlou, A. H. Contribution of human embryonic stem cells to mouse blastocysts. Dev. Biol. 295, 90\u2013102 (2006).","journal-title":"Dev. Biol."},{"key":"565_CR6","doi-asserted-by":"publisher","first-page":"1292","DOI":"10.1038\/nature08534","volume":"461","author":"S Bao","year":"2009","unstructured":"Bao, S. et al. Epigenetic reversion of post-implantation epiblast to pluripotent embryonic stem cells. Nature 461, 1292\u20131295 (2009).","journal-title":"Nature"},{"key":"565_CR7","doi-asserted-by":"publisher","first-page":"103","DOI":"10.1016\/j.stemcr.2014.12.003","volume":"4","author":"H Murayama","year":"2015","unstructured":"Murayama, H. et al. Successful reprogramming of epiblast stem cells by blocking nuclear localization of beta-catenin. Stem Cell Rep. 4, 103\u2013113 (2015).","journal-title":"Stem Cell Rep."},{"key":"565_CR8","doi-asserted-by":"publisher","first-page":"519","DOI":"10.1038\/nature06968","volume":"453","author":"QL Ying","year":"2008","unstructured":"Ying, Q. L. et al. The ground state of embryonic stem cell self-renewal. Nature 453, 519\u2013523 (2008).","journal-title":"Nature"},{"key":"565_CR9","doi-asserted-by":"publisher","first-page":"282","DOI":"10.1038\/nature12745","volume":"504","author":"O Gafni","year":"2013","unstructured":"Gafni, O. et al. Derivation of novel human ground state naive pluripotent stem cells. Nature 504, 282\u2013286 (2013).","journal-title":"Nature"},{"key":"565_CR10","doi-asserted-by":"publisher","first-page":"663","DOI":"10.1016\/j.stem.2013.11.015","volume":"13","author":"YS Chan","year":"2013","unstructured":"Chan, Y. S. et al. Induction of a human pluripotent state with distinct regulatory circuitry that resembles preimplantation epiblast. Cell Stem Cell 13, 663\u2013675 (2013).","journal-title":"Cell Stem Cell"},{"key":"565_CR11","doi-asserted-by":"publisher","first-page":"524","DOI":"10.1016\/j.stem.2014.09.003","volume":"15","author":"TW Theunissen","year":"2014","unstructured":"Theunissen, T. W. et al. Systematic identification of culture conditions for induction and maintenance of naive human pluripotency. Cell Stem Cell 15, 524\u2013526 (2014).","journal-title":"Cell Stem Cell"},{"key":"565_CR12","doi-asserted-by":"publisher","first-page":"4484","DOI":"10.1073\/pnas.1319738111","volume":"111","author":"CB Ware","year":"2014","unstructured":"Ware, C. B. et al. Derivation of naive human embryonic stem cells. Proc. Natl Acad. Sci. USA 111, 4484\u20134489 (2014).","journal-title":"Proc. Natl Acad. Sci. USA"},{"key":"565_CR13","doi-asserted-by":"publisher","first-page":"473","DOI":"10.1016\/j.cell.2016.12.036","volume":"168","author":"J Wu","year":"2017","unstructured":"Wu, J. et al. Interspecies chimerism with mammalian pluripotent stem cells. Cell 168, 473\u2013486 e415 (2017).","journal-title":"Cell"},{"key":"565_CR14","doi-asserted-by":"publisher","first-page":"243","DOI":"10.1016\/j.cell.2017.02.005","volume":"169","author":"Y Yang","year":"2017","unstructured":"Yang, Y. et al. Derivation of pluripotent stem cells with in vivo embryonic and extraembryonic potency. Cell 169, 243\u2013257 e225 (2017).","journal-title":"Cell"},{"key":"565_CR15","doi-asserted-by":"publisher","first-page":"155","DOI":"10.1038\/nrm.2015.28","volume":"17","author":"L Weinberger","year":"2016","unstructured":"Weinberger, L., Ayyash, M., Novershtern, N. & Hanna, J. H. Dynamic stem cell states: naive to primed pluripotency in rodents and humans. Nat. Rev. Mol. Cell Biol. 17, 155\u2013169 (2016).","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"565_CR16","doi-asserted-by":"publisher","first-page":"301","DOI":"10.1016\/j.stem.2016.02.012","volume":"18","author":"Y Wang","year":"2016","unstructured":"Wang, Y. & Gao, S. Human naive embryonic stem cells: how full is the glass? Cell Stem Cell 18, 301\u2013303 (2016).","journal-title":"Cell Stem Cell"},{"key":"565_CR17","doi-asserted-by":"publisher","first-page":"eaaz0298","DOI":"10.1126\/sciadv.aaz0298","volume":"6","author":"Z Hu","year":"2020","unstructured":"Hu, Z. et al. Transient inhibition of mTOR in human pluripotent stem cells enables robust formation of mouse-human chimeric embryos. Sci. Adv. 6, eaaz0298 (2020).","journal-title":"Sci. Adv."},{"key":"565_CR18","doi-asserted-by":"publisher","first-page":"535","DOI":"10.1016\/j.stem.2010.05.003","volume":"6","author":"C Buecker","year":"2010","unstructured":"Buecker, C. et al. A murine ESC-like state facilitates transgenesis and homologous recombination in human pluripotent stem cells. Cell Stem Cell 6, 535\u2013546 (2010).","journal-title":"Cell Stem Cell"},{"key":"565_CR19","doi-asserted-by":"publisher","first-page":"9222","DOI":"10.1073\/pnas.1004584107","volume":"107","author":"J Hanna","year":"2010","unstructured":"Hanna, J. et al. Human embryonic stem cells with biological and epigenetic characteristics similar to those of mouse ESCs. Proc. Natl Acad. Sci. USA 107, 9222\u20139227 (2010).","journal-title":"Proc. Natl Acad. Sci. USA"},{"key":"565_CR20","doi-asserted-by":"publisher","first-page":"18283","DOI":"10.1073\/pnas.1100893108","volume":"108","author":"W Wang","year":"2011","unstructured":"Wang, W. et al. Rapid and efficient reprogramming of somatic cells to induced pluripotent stem cells by retinoic acid receptor gamma and liver receptor homolog 1. Proc. Natl Acad. Sci. USA 108, 18283\u201318288 (2011).","journal-title":"Proc. Natl Acad. Sci. USA"},{"key":"565_CR21","doi-asserted-by":"publisher","first-page":"2591","DOI":"10.1089\/scd.2015.0079","volume":"24","author":"Z Hu","year":"2015","unstructured":"Hu, Z. et al. Generation of naivetropic induced pluripotent stem cells from Parkinson\u2019s disease patients for high-efficiency genetic manipulation and disease modeling. Stem Cells Dev. 24, 2591\u20132604 (2015).","journal-title":"Stem Cells Dev."},{"key":"565_CR22","doi-asserted-by":"publisher","first-page":"1254","DOI":"10.1016\/j.cell.2014.08.029","volume":"158","author":"Y Takashima","year":"2014","unstructured":"Takashima, Y. et al. Resetting transcription factor control circuitry toward ground-state pluripotency in human. Cell 158, 1254\u20131269 (2014).","journal-title":"Cell"},{"key":"565_CR23","doi-asserted-by":"publisher","first-page":"410","DOI":"10.1016\/j.stem.2014.09.014","volume":"15","author":"K Huang","year":"2014","unstructured":"Huang, K., Maruyama, T. & Fan, G. The naive state of human pluripotent stem cells: a synthesis of stem cell and preimplantation embryo transcriptome analyses. Cell Stem Cell 15, 410\u2013415 (2014).","journal-title":"Cell Stem Cell"},{"key":"565_CR24","doi-asserted-by":"publisher","first-page":"502","DOI":"10.1016\/j.stem.2016.06.011","volume":"19","author":"TW Theunissen","year":"2016","unstructured":"Theunissen, T. W. et al. Molecular criteria for defining the naive human pluripotent state. Cell Stem Cell 19, 502\u2013515 (2016).","journal-title":"Cell Stem Cell"},{"key":"565_CR25","doi-asserted-by":"publisher","first-page":"393","DOI":"10.1038\/nature24052","volume":"550","author":"J Yang","year":"2017","unstructured":"Yang, J. et al. Establishment of mouse expanded potential stem cells. Nature 550, 393\u2013397 (2017).","journal-title":"Nature"},{"key":"565_CR26","doi-asserted-by":"publisher","first-page":"687","DOI":"10.1038\/s41556-019-0333-2","volume":"21","author":"X Gao","year":"2019","unstructured":"Gao, X. et al. Establishment of porcine and human expanded potential stem cells. Nat. Cell Biol. 21, 687\u2013699 (2019).","journal-title":"Nat. Cell Biol."},{"key":"565_CR27","doi-asserted-by":"publisher","first-page":"335","DOI":"10.1016\/j.cell.2013.03.012","volume":"153","author":"J Betschinger","year":"2013","unstructured":"Betschinger, J. et al. Exit from pluripotency is gated by intracellular redistribution of the bHLH transcription factor Tfe3. Cell 153, 335\u2013347 (2013).","journal-title":"Cell"},{"key":"565_CR28","doi-asserted-by":"crossref","unstructured":"Raben, N. & Puertollano, R. TFEB and TFE3: linking lysosomes to cellular adaptation to stress. Annu. Rev. Cell Dev. Biol (2016).","DOI":"10.1146\/annurev-cellbio-111315-125407"},{"key":"565_CR29","doi-asserted-by":"publisher","first-page":"ra9","DOI":"10.1126\/scisignal.2004754","volume":"7","author":"JA Martina","year":"2014","unstructured":"Martina, J. A. et al. The nutrient-responsive transcription factor TFE3 promotes autophagy, lysosomal biogenesis, and clearance of cellular debris. Sci. Signal 7, ra9 (2014).","journal-title":"Sci. Signal"},{"key":"565_CR30","doi-asserted-by":"publisher","first-page":"1570","DOI":"10.1073\/pnas.1525518113","volume":"113","author":"MA Cohen","year":"2016","unstructured":"Cohen, M. A. et al. Human neural crest cells contribute to coat pigmentation in interspecies chimeras after in utero injection into mouse embryos. Proc. Natl Acad. Sci. USA 113, 1570\u20131575 (2016).","journal-title":"Proc. Natl Acad. Sci. USA"},{"key":"565_CR31","doi-asserted-by":"publisher","first-page":"13","DOI":"10.1101\/gr.6858507","volume":"18","author":"DM Stults","year":"2008","unstructured":"Stults, D. M., Killen, M. W., Pierce, H. H. & Pierce, A. J. Genomic architecture and inheritance of human ribosomal RNA gene clusters. Genome Res. 18, 13\u201318 (2008).","journal-title":"Genome Res."},{"key":"565_CR32","first-page":"419","volume":"38","author":"IL Gonzalez","year":"1986","unstructured":"Gonzalez, I. L. & Schmickel, R. D. The human 18S ribosomal RNA gene: evolution and stability. Am. J. Hum. Genet 38, 419\u2013427 (1986).","journal-title":"Am. J. Hum. Genet"},{"key":"565_CR33","doi-asserted-by":"publisher","first-page":"787","DOI":"10.1016\/j.cell.2010.07.039","volume":"142","author":"T Kobayashi","year":"2010","unstructured":"Kobayashi, T. et al. Generation of rat pancreas in mouse by interspecific blastocyst injection of pluripotent stem cells. Cell 142, 787\u2013799 (2010).","journal-title":"Cell"},{"key":"565_CR34","doi-asserted-by":"publisher","first-page":"191","DOI":"10.1038\/nature21070","volume":"542","author":"T Yamaguchi","year":"2017","unstructured":"Yamaguchi, T. et al. Interspecies organogenesis generates autologous functional islets. Nature 542, 191\u2013196 (2017).","journal-title":"Nature"},{"key":"565_CR35","doi-asserted-by":"publisher","first-page":"51","DOI":"10.1038\/nature20573","volume":"540","author":"J Wu","year":"2016","unstructured":"Wu, J. et al. Stem cells and interspecies chimaeras. Nature 540, 51\u201359 (2016).","journal-title":"Nature"},{"key":"565_CR36","doi-asserted-by":"publisher","first-page":"257","DOI":"10.1016\/j.stem.2018.11.021","volume":"24","author":"F Villegas","year":"2019","unstructured":"Villegas, F. et al. Lysosomal signaling licenses embryonic stem cell differentiation via inactivation of Tfe3. Cell Stem Cell 24, 257\u2013270 (2019).","journal-title":"Cell Stem Cell"},{"key":"565_CR37","doi-asserted-by":"publisher","first-page":"2485","DOI":"10.1073\/pnas.1416878112","volume":"112","author":"JG Gibbons","year":"2015","unstructured":"Gibbons, J. G., Branco, A. T., Godinho, S. A., Yu, S. & Lemos, B. Concerted copy number variation balances ribosomal DNA dosage in human and mouse genomes. Proc. Natl Acad. Sci. USA 112, 2485\u20132490 (2015).","journal-title":"Proc. Natl Acad. Sci. USA"},{"key":"565_CR38","doi-asserted-by":"publisher","first-page":"747","DOI":"10.1128\/CMR.00114-16","volume":"30","author":"KB Waites","year":"2017","unstructured":"Waites, K. B., Xiao, L., Liu, Y., Balish, M. F. & Atkinson, T. P. Mycoplasma pneumoniae from the respiratory tract and beyond. Clin. Microbiol. Rev. 30, 747\u2013809 (2017).","journal-title":"Clin. Microbiol. Rev."},{"key":"565_CR39","unstructured":"Nagy, A. Manipulating the Mouse Embryo: A Laboratory Manual 3rd edn (Cold Spring Harbor Laboratory Press, 2003)."},{"key":"565_CR40","doi-asserted-by":"publisher","first-page":"93","DOI":"10.1007\/978-1-61779-080-5_8","volume":"731","author":"CC Uphoff","year":"2011","unstructured":"Uphoff, C. C. & Drexler, H. G. Detecting mycoplasma contamination in cell cultures by polymerase chain reaction. Methods Mol. Biol. 731, 93\u2013103 (2011).","journal-title":"Methods Mol. Biol."},{"key":"565_CR41","doi-asserted-by":"publisher","first-page":"3047","DOI":"10.1093\/bioinformatics\/btw354","volume":"32","author":"P Ewels","year":"2016","unstructured":"Ewels, P., Magnusson, M., Lundin, S. & Kaller, M. MultiQC: summarize analysis results for multiple tools and samples in a single report. Bioinformatics 32, 3047\u20133048 (2016).","journal-title":"Bioinformatics"},{"key":"565_CR42","doi-asserted-by":"publisher","first-page":"10","DOI":"10.14806\/ej.17.1.200","volume":"17","author":"M Martin","year":"2011","unstructured":"Martin, M. Cutadapt removes adapter sequences from high-throughput sequencing reads. EMBnet. J. 17, 10\u201312 (2011).","journal-title":"EMBnet. J."},{"key":"565_CR43","doi-asserted-by":"publisher","first-page":"e2584","DOI":"10.7717\/peerj.2584","volume":"4","author":"T Rognes","year":"2016","unstructured":"Rognes, T., Flouri, T., Nichols, B., Quince, C. & Mahe, F. VSEARCH: a versatile open source tool for metagenomics. PeerJ 4, e2584 (2016).","journal-title":"PeerJ"},{"key":"565_CR44","doi-asserted-by":"publisher","first-page":"559","DOI":"10.1038\/nprot.2014.030","volume":"9","author":"A Czechanski","year":"2014","unstructured":"Czechanski, A. et al. Derivation and characterization of mouse embryonic stem cells from permissive and nonpermissive strains. Nat. Protoc. 9, 559\u2013574 (2014).","journal-title":"Nat. Protoc."},{"key":"565_CR45","doi-asserted-by":"publisher","first-page":"409","DOI":"10.1038\/nmeth.1591","volume":"8","author":"K Okita","year":"2011","unstructured":"Okita, K. et al. A more efficient method to generate integration-free human iPS cells. Nat. Methods 8, 409\u2013412 (2011).","journal-title":"Nat. Methods"},{"key":"565_CR46","doi-asserted-by":"publisher","first-page":"418","DOI":"10.1038\/nprot.2009.231","volume":"5","author":"K Okita","year":"2010","unstructured":"Okita, K., Hong, H., Takahashi, K. & Yamanaka, S. Generation of mouse-induced pluripotent stem cells with plasmid vectors. Nat. Protoc. 5, 418\u2013428 (2010).","journal-title":"Nat. Protoc."}],"container-title":["Nature Protocols"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41596-021-00565-7.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41596-021-00565-7","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41596-021-00565-7.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,8,4]],"date-time":"2021-08-04T16:12:08Z","timestamp":1628093528000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41596-021-00565-7"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,7,2]]},"references-count":46,"journal-issue":{"issue":"8","published-print":{"date-parts":[[2021,8]]}},"alternative-id":["565"],"URL":"https:\/\/doi.org\/10.1038\/s41596-021-00565-7","relation":{},"ISSN":["1754-2189","1750-2799"],"issn-type":[{"value":"1754-2189","type":"print"},{"value":"1750-2799","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,7,2]]},"assertion":[{"value":"1 November 2020","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"29 April 2021","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"2 July 2021","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"J.F. is a cofounder of Vitropy, LLC and ASDDR, LLC, and has a patent application regarding this work (no. 16\/346534). All other authors declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}]}}