@article{42df6eef7863465a8e15cca7da41035f,
title = "Generation of Blastocyst-like Structures from Mouse Embryonic and Adult Cell Cultures",
abstract = "A single mouse blastomere from an embryo until the 8-cell stage can generate an entire blastocyst. Whether laboratory-cultured cells retain a similar generative capacity remains unknown. Starting from a single stem cell type, extended pluripotent stem (EPS) cells, we established a 3D differentiation system that enabled the generation of blastocyst-like structures (EPS-blastoids) through lineage segregation and self-organization. EPS-blastoids resembled blastocysts in morphology and cell-lineage allocation and recapitulated key morphogenetic events during preimplantation and early postimplantation development in vitro. Upon transfer, some EPS-blastoids underwent implantation, induced decidualization, and generated live, albeit disorganized, tissues in utero. Single-cell and bulk RNA-sequencing analysis revealed that EPS-blastoids contained all three blastocyst cell lineages and shared transcriptional similarity with natural blastocysts. We also provide proof of concept that EPS-blastoids can be generated from adult cells via cellular reprogramming. EPS-blastoids provide a unique platform for studying early embryogenesis and pave the way to creating viable synthetic embryos by using cultured cells.",
keywords = "EPS cells, EPS-blastoid, blastocyst, blastoids, extended pluripotent stem cells, implantation, reprogramming, totipotent",
author = "Ronghui Li and Cuiqing Zhong and Yang Yu and Haisong Liu and Masahiro Sakurai and Leqian Yu and Zheying Min and Lei Shi and Yulei Wei and Yuta Takahashi and Liao, {Hsin Kai} and Jie Qiao and Hongkui Deng and Estrella Nu{\~n}ez-Delicado and {Rodriguez Esteban}, Concepcion and Jun Wu and {Izpisua Belmonte}, {Juan Carlos}",
note = "Funding Information: We thank Travis Berggren for his continuous advice and guidance. We thank Dr. Fumihiro for the B6N-22 mESC line. We are thankful to Dr. Azim Surani for the female X-GFP mEpiSC. We are grateful for technical assistance from C. O'Connor and C. Fitzpatrick from the Salk Institute Flow Cytometry Core Facility for FACS sorting, U. Manor and T. Zhang from the Waitt Advanced Biophotonics Core for confocal imaging, and N. Hah and G. Chou from the Salk Sequencing Core for single-cell RNA-seq experiments. We are also thankful to all the GEL-B lab members for helpful discussion and constructive criticism and M. Schwarz and P. Schwarz for administrative assistance. We would also like to express our gratitude to Dave O'Keefe for his contribution toward manuscript preparation. C.Z. was supported by the Larry L. Hillblom Foundation, Paul F. Glenn Foundation, and Salk Women & Science Special Award. This work was also supported by the National Key R&D Program of China (2016YFC1000601 to Y.Y.). L.Y. was partially supported by a trainee fellowship from the Hamon Center for Regenerative Science & Medicine. J.W. was a Virginia Murchison Linthicum Scholar in Medical Research and funded by Cancer Prevention & Research Institute of Texas (CPRIT). This project was supported by the G. Harold and Leila Y. Mathers Charitable Foundation, the Moxie Foundation, the Leona M. and Harry B. Helmsley Charitable Trust (2012-PG-MED002), the Hewitt Foundation, the NIH (5 DP1 DK113616), and Universidad Cato?lica San Antonio de Murcia. R.L. J.W. and J.C.I.B. conceived the study, designed experiments, and interpreted results. R.L. performed experiments, collected and analyzed the data, and wrote the manuscript. C.Z. performed EPS-blastoid induction experiments and immunofluorescence staining. Y.Y. conducted EPS-blastoid transfers and microinjections, collected samples for RNA-seq, and analyzed RNA-seq data. H.L. performed chemical screening, immunofluorescence staining, and optimization of single-cell RNA-seq procedures. M.S. and C.R.E. performed EPS-blastoid transfers and microinjection. L.Y. performed XEN derivations and characterization. Z.M. dissected embryos out of decidua. L.S. performed single-cell RNA-seq data analysis. Y.W. performed XEN cell microinjections and immunofluorescence staining. Y.T. collected natural embryos. H.-K.L. prepared lentiviral particle. J.Q. H.D. and E.N.D. supervised the study and interpreted the results. J.W. and J.C.I.B. supervised the study and wrote the manuscript with comments from all authors. The authors declare no competing interests. Funding Information: We thank Travis Berggren for his continuous advice and guidance. We thank Dr. Fumihiro for the B6N-22 mESC line. We are thankful to Dr. Azim Surani for the female X-GFP mEpiSC. We are grateful for technical assistance from C. O{\textquoteright}Connor and C. Fitzpatrick from the Salk Institute Flow Cytometry Core Facility for FACS sorting, U. Manor and T. Zhang from the Waitt Advanced Biophotonics Core for confocal imaging, and N. Hah and G. Chou from the Salk Sequencing Core for single-cell RNA-seq experiments. We are also thankful to all the GEL-B lab members for helpful discussion and constructive criticism and M. Schwarz and P. Schwarz for administrative assistance. We would also like to express our gratitude to Dave O{\textquoteright}Keefe for his contribution toward manuscript preparation. C.Z. was supported by the Larry L. Hillblom Foundation , Paul F. Glenn Foundation , and Salk Women & Science Special Award. This work was also supported by the National Key R&D Program of China ( 2016YFC1000601 to Y.Y.). L.Y. was partially supported by a trainee fellowship from the Hamon Center for Regenerative Science & Medicine . J.W. was a Virginia Murchison Linthicum Scholar in Medical Research and funded by Cancer Prevention & Research Institute of Texas (CPRIT). This project was supported by the G. Harold and Leila Y. Mathers Charitable Foundation , the Moxie Foundation , the Leona M. and Harry B. Helmsley Charitable Trust ( 2012-PG-MED002 ), the Hewitt Foundation , the NIH ( 5 DP1 DK113616 ), and Universidad Cat{\'o}lica San Antonio de Murcia . Publisher Copyright: {\textcopyright} 2019 Elsevier Inc.",
year = "2019",
month = oct,
day = "17",
doi = "10.1016/j.cell.2019.09.029",
language = "English (US)",
volume = "179",
pages = "687--702.e18",
journal = "Cell",
issn = "0092-8674",
publisher = "Cell Press",
number = "3",
}