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Derivation of human extended pluripotent stem cells in feeder-free condition

Ran Zheng, Ting Geng, Dan-Ya Wu, Tianzhe Zhang, Hai-Nan He, Liyan Wang, Haining Du, Donghui Zhang, Yi-Liang Miao, View ORCID ProfileWei Jiang
doi: https://doi.org/10.1101/2020.10.18.344259
Ran Zheng
1Department of Biological Repositories, Frontier Science Center for Immunology and Metabolism, Medical Research Institute, Zhongnan Hospital of Wuhan University, Wuhan University, Wuhan 430071, China
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Ting Geng
1Department of Biological Repositories, Frontier Science Center for Immunology and Metabolism, Medical Research Institute, Zhongnan Hospital of Wuhan University, Wuhan University, Wuhan 430071, China
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Dan-Ya Wu
2Institute of Stem Cell and Regenerative Biology, College of Animal Science and Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China
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Tianzhe Zhang
1Department of Biological Repositories, Frontier Science Center for Immunology and Metabolism, Medical Research Institute, Zhongnan Hospital of Wuhan University, Wuhan University, Wuhan 430071, China
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Hai-Nan He
2Institute of Stem Cell and Regenerative Biology, College of Animal Science and Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China
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Liyan Wang
2Institute of Stem Cell and Regenerative Biology, College of Animal Science and Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China
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Haining Du
4Hubei Key Laboratory of Cell Homeostasis, College of Life Sciences, Wuhan University, Wuhan 430071, China
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Donghui Zhang
3State Key Laboratory of Biocatalysis and Enzyme Engineering, School of Life Science, Hubei University, Wuhan 430062, China
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  • For correspondence: jiangw.mri@whu.edu.cn miaoyl@mail.hzau.edu.cn dongh.zhang@hubu.edu.cn
Yi-Liang Miao
2Institute of Stem Cell and Regenerative Biology, College of Animal Science and Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China
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  • For correspondence: jiangw.mri@whu.edu.cn miaoyl@mail.hzau.edu.cn dongh.zhang@hubu.edu.cn
Wei Jiang
1Department of Biological Repositories, Frontier Science Center for Immunology and Metabolism, Medical Research Institute, Zhongnan Hospital of Wuhan University, Wuhan University, Wuhan 430071, China
5Human Genetics Resource Preservation Center of Wuhan University, Wuhan 430071, China
6Hubei Provincial Key Laboratory of Developmentally Originated Disease, Wuhan 430071, China
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  • ORCID record for Wei Jiang
  • For correspondence: jiangw.mri@whu.edu.cn miaoyl@mail.hzau.edu.cn dongh.zhang@hubu.edu.cn
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SUMMARY

Human extended pluripotent stem cells (EPSCs), with bidirectional chimeric ability to contribute to both embryonic and extra-embryonic lineages, can be obtained and maintained by converting embryonic stem cells (ESCs) using chemicals. However, the transition system is based on inactivated mouse fibroblast, which greatly hinders the mechanistic studies of extended pluripotency and further applications. Here we reported a Matrigel-based feeder-free method to convert human ESCs and iPSCs into EPSCs and demonstrated the extended pluripotency in terms of molecular features, chimeric ability, and transcriptome. We further improved the protocol by applying chemicals targeting glycolysis and histone methyltransferase. Altogether, our data established a feeder-free system to generate human EPSCs and provided additional insights into the acquisition of extended pluripotency.

Competing Interest Statement

The authors have declared no competing interest.

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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Posted October 18, 2020.
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Derivation of human extended pluripotent stem cells in feeder-free condition
Ran Zheng, Ting Geng, Dan-Ya Wu, Tianzhe Zhang, Hai-Nan He, Liyan Wang, Haining Du, Donghui Zhang, Yi-Liang Miao, Wei Jiang
bioRxiv 2020.10.18.344259; doi: https://doi.org/10.1101/2020.10.18.344259
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Derivation of human extended pluripotent stem cells in feeder-free condition
Ran Zheng, Ting Geng, Dan-Ya Wu, Tianzhe Zhang, Hai-Nan He, Liyan Wang, Haining Du, Donghui Zhang, Yi-Liang Miao, Wei Jiang
bioRxiv 2020.10.18.344259; doi: https://doi.org/10.1101/2020.10.18.344259

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