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Generation of human blastocyst-like structures from pluripotent stem cells

Yong Fan, Zhe-Ying Min, Samhan Alsolami, Zheng-Lai Ma, Ke Zhong, Wen-Di Pei, Pu-Yao Zhang, Xiang-Jin Kang, Ying-Ying Zhang, Hai-Ying Zhu, Jie Qiao, Mo Li, Yang Yu
doi: https://doi.org/10.1101/2021.03.09.434313
Yong Fan
1Department of Gynecology and Obstetrics, Key Laboratory for Major Obstetric Diseases of Guangdong Province, Key Laboratory of Reproduction and Genetics of Guangdong Higher Education Institutes, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, 510150, China
4Stem Cell Research Center, Peking University Third Hospital, Beijing 100191, China
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  • For correspondence: yuyang5012@hotmail.com mo.li@kaust.edu.sa yongfan011@gzhmu.edu.cn
Zhe-Ying Min
2Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology and Key Laboratory of Assisted Reproduction, Ministry of Education, Center of Reproductive Medicine, Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing, 100191, China
4Stem Cell Research Center, Peking University Third Hospital, Beijing 100191, China
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Samhan Alsolami
3Biological and Environmental Sciences and Engineering Division, King Abdullah University of Science and Technology (KAUST), Thuwal 23955-6900, Kingdom of Saudi Arabia
4Stem Cell Research Center, Peking University Third Hospital, Beijing 100191, China
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Zheng-Lai Ma
1Department of Gynecology and Obstetrics, Key Laboratory for Major Obstetric Diseases of Guangdong Province, Key Laboratory of Reproduction and Genetics of Guangdong Higher Education Institutes, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, 510150, China
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Ke Zhong
1Department of Gynecology and Obstetrics, Key Laboratory for Major Obstetric Diseases of Guangdong Province, Key Laboratory of Reproduction and Genetics of Guangdong Higher Education Institutes, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, 510150, China
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Wen-Di Pei
2Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology and Key Laboratory of Assisted Reproduction, Ministry of Education, Center of Reproductive Medicine, Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing, 100191, China
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Pu-Yao Zhang
2Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology and Key Laboratory of Assisted Reproduction, Ministry of Education, Center of Reproductive Medicine, Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing, 100191, China
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Xiang-Jin Kang
1Department of Gynecology and Obstetrics, Key Laboratory for Major Obstetric Diseases of Guangdong Province, Key Laboratory of Reproduction and Genetics of Guangdong Higher Education Institutes, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, 510150, China
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Ying-Ying Zhang
1Department of Gynecology and Obstetrics, Key Laboratory for Major Obstetric Diseases of Guangdong Province, Key Laboratory of Reproduction and Genetics of Guangdong Higher Education Institutes, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, 510150, China
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Hai-Ying Zhu
1Department of Gynecology and Obstetrics, Key Laboratory for Major Obstetric Diseases of Guangdong Province, Key Laboratory of Reproduction and Genetics of Guangdong Higher Education Institutes, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, 510150, China
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Jie Qiao
2Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology and Key Laboratory of Assisted Reproduction, Ministry of Education, Center of Reproductive Medicine, Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing, 100191, China
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Mo Li
3Biological and Environmental Sciences and Engineering Division, King Abdullah University of Science and Technology (KAUST), Thuwal 23955-6900, Kingdom of Saudi Arabia
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  • For correspondence: yuyang5012@hotmail.com mo.li@kaust.edu.sa yongfan011@gzhmu.edu.cn
Yang Yu
1Department of Gynecology and Obstetrics, Key Laboratory for Major Obstetric Diseases of Guangdong Province, Key Laboratory of Reproduction and Genetics of Guangdong Higher Education Institutes, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, 510150, China
2Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology and Key Laboratory of Assisted Reproduction, Ministry of Education, Center of Reproductive Medicine, Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing, 100191, China
4Stem Cell Research Center, Peking University Third Hospital, Beijing 100191, China
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  • For correspondence: yuyang5012@hotmail.com mo.li@kaust.edu.sa yongfan011@gzhmu.edu.cn
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Abstract

Human blastocysts are comprised of the first three cell lineages of the embryo: trophectoderm, epiblast, and primitive endoderm, all of which are essential for early development and organ formation1,2. However, due to ethical concerns and restricted access to human blastocysts, we lack a comprehensive understanding of early human embryogenesis. To bridge this knowledge gap, we need a reliable model system that recapitulates early stages of human embryogenesis. Here we report a ∼three-dimensional (3D), two-step induction protocol for generating blastocyst-like structures (EPS-blastoids) from human extended pluripotent stem (EPS) cells. Morphological and single-cell transcriptomic analyses revealed that EPS-blastoids contain key cell lineages and are transcriptionally similar to human blastocysts. Furthermore, EPS-blastoids also exhibited the developmental potential to undergo post-implantation morphogenesis in vitro to form structures with a cellular composition and transcriptome signature similar to human embryos that had been cultured in vitro for 8 or 10 days. In conclusion, human EPS-blastoids provide a new experimental platform for studying early developmental stages of the human embryo.

Highlights A method for generating human blastoids from EPS cells.

Human blastoids resemble blastocysts in terms of morphology and cell lineage composition.

Single-cell transcriptomic analyses reveal EPI, PE, and TE cell lineages in human blastoids.

Human blastoids mimic in vitro the morphogenetic events of pre- and early post-implantation stages.

Competing Interest Statement

The authors have declared no competing interest.

Footnotes

  • Lead contact: Y.Y.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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Generation of human blastocyst-like structures from pluripotent stem cells
Yong Fan, Zhe-Ying Min, Samhan Alsolami, Zheng-Lai Ma, Ke Zhong, Wen-Di Pei, Pu-Yao Zhang, Xiang-Jin Kang, Ying-Ying Zhang, Hai-Ying Zhu, Jie Qiao, Mo Li, Yang Yu
bioRxiv 2021.03.09.434313; doi: https://doi.org/10.1101/2021.03.09.434313
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Generation of human blastocyst-like structures from pluripotent stem cells
Yong Fan, Zhe-Ying Min, Samhan Alsolami, Zheng-Lai Ma, Ke Zhong, Wen-Di Pei, Pu-Yao Zhang, Xiang-Jin Kang, Ying-Ying Zhang, Hai-Ying Zhu, Jie Qiao, Mo Li, Yang Yu
bioRxiv 2021.03.09.434313; doi: https://doi.org/10.1101/2021.03.09.434313

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