A Universal 6iL/E4 Culture System for Deriving and Maintaining Embryonic Stem Cells Across Mammalian Species Article Swipe
Duo Wang
,
Ming Hao
,
Dongshan Yang
,
Li‐Kuang Tsai
,
Zhuying Wei
,
Giovanna Nascimento Scatolin
,
Xiukun Wang
,
King‐Wai Yau
,
Litao Tao
,
Xinyi Tong
,
Shuling Wang
,
Kai‐Xuan Shi
,
Denis Evseenko
,
Ben Van Handel
,
Bingjing Zhang
,
Yinjuan Wang
,
Rajan Iyyappan
,
Oscar Alejandro Ojeda-Rojas
,
Guang Hu
,
Lynda K. McGinnis
,
Richard J. Paulson
,
Daniel B. McKim
,
Xiangbo Kong
,
Xiaofeng Xia
,
Jifeng Zhang
,
Y. Eugene Chen
,
Jie Xu
,
Zongliang Jiang
,
Qi‐Long Ying
·
YOU?
·
· 2025
· Open Access
·
· DOI: https://doi.org/10.1101/2025.05.20.654948
YOU?
·
· 2025
· Open Access
·
· DOI: https://doi.org/10.1101/2025.05.20.654948
SUMMARY The derivation of authentic embryonic stem cells (ESCs) from diverse mammalian species offers valuable opportunities for advancing regenerative medicine, studying developmental biology, and enabling species conservation. Here, we report the development of a robust, serum-free culture system, termed 6iL/E4 that enables the derivation and long-term self-renewal of ESCs from multiple mammalian species, including mouse, rat, bovine, rabbit, and human. Using systematic signaling pathway analysis, we identified key regulators—including GSK3α, STAT3, PDGFR, BRAF, and LATS—critical for ESC maintenance across species. Additionally, inducible expression of KLF2 and NANOG enhances the naive pluripotency and chimeric potential of bovine ESCs. The E4 medium also supports stable ESC growth while minimizing lineage bias. These findings reveal conserved principles underlying ESC self-renewal across divergent mammalian species and provide a universal platform for cross-species stem cell research, disease modeling, and biotechnology applications. In Brief Wang et al. developed 6iL/E4, a serum-free system sustaining ESCs from mouse, rat, bovine, rabbit, and human. These findings reveal conserved fundamental mechanisms governing ESC self-renewal across diverse mammalian species. Highlights Developed 6iL/E4 system for ESC derivation across five mammalian species. PDGFR signaling inhibition as critical for ESC derivation across species. E4 medium improves ESC maintenance and avoids neural bias of traditional N2B27. Inducible KLF2/NANOG enhances naive pluripotency and chimera formation in bovine.
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- Type
- preprint
- Language
- en
- Landing Page
- https://doi.org/10.1101/2025.05.20.654948
- https://www.biorxiv.org/content/biorxiv/early/2025/05/21/2025.05.20.654948.full.pdf
- OA Status
- green
- Cited By
- 1
- References
- 61
- Related Works
- 10
- OpenAlex ID
- https://openalex.org/W4410556446
All OpenAlex metadata
Raw OpenAlex JSON
- OpenAlex ID
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https://openalex.org/W4410556446Canonical identifier for this work in OpenAlex
- DOI
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https://doi.org/10.1101/2025.05.20.654948Digital Object Identifier
- Title
-
A Universal 6iL/E4 Culture System for Deriving and Maintaining Embryonic Stem Cells Across Mammalian SpeciesWork title
- Type
-
preprintOpenAlex work type
- Language
-
enPrimary language
- Publication year
-
2025Year of publication
- Publication date
-
2025-05-21Full publication date if available
- Authors
-
Duo Wang, Ming Hao, Dongshan Yang, Li‐Kuang Tsai, Zhuying Wei, Giovanna Nascimento Scatolin, Xiukun Wang, King‐Wai Yau, Litao Tao, Xinyi Tong, Shuling Wang, Kai‐Xuan Shi, Denis Evseenko, Ben Van Handel, Bingjing Zhang, Yinjuan Wang, Rajan Iyyappan, Oscar Alejandro Ojeda-Rojas, Guang Hu, Lynda K. McGinnis, Richard J. Paulson, Daniel B. McKim, Xiangbo Kong, Xiaofeng Xia, Jifeng Zhang, Y. Eugene Chen, Jie Xu, Zongliang Jiang, Qi‐Long YingList of authors in order
- Landing page
-
https://doi.org/10.1101/2025.05.20.654948Publisher landing page
- PDF URL
-
https://www.biorxiv.org/content/biorxiv/early/2025/05/21/2025.05.20.654948.full.pdfDirect link to full text PDF
- Open access
-
YesWhether a free full text is available
- OA status
-
greenOpen access status per OpenAlex
- OA URL
-
https://www.biorxiv.org/content/biorxiv/early/2025/05/21/2025.05.20.654948.full.pdfDirect OA link when available
- Concepts
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Embryonic stem cell, Stem cell, Biology, Cell biology, Computational biology, Evolutionary biology, Genetics, GeneTop concepts (fields/topics) attached by OpenAlex
- Cited by
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1Total citation count in OpenAlex
- Citations by year (recent)
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2025: 1Per-year citation counts (last 5 years)
- References (count)
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61Number of works referenced by this work
- Related works (count)
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10Other works algorithmically related by OpenAlex
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