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Volume 15
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10.3390/cells15100954
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This is an early access version, the complete PDF, HTML, and XML versions will be available soon.
Article

The Trilineage Coexistence Observed During the Differentiation of Porcine EPSCs

by
Lihua Zhao
1,2,
Yanglin Chen
1,3,
Xiyun Guo
2,
Meng Zhou
2,
Tianxu Guo
2,
Junjun Ma
2,
Manling Zhang
2,
Linxin Cheng
2,
Jinbo Yu
2,
Yu Zhang
2,
Guang Yang
2,
Rongfeng Li
2,4,5,* and
Xihe Li
1,3,5,*
1
The State Key Laboratory of Reproductive Regulation and Breeding of Grassland Livestock, College of Life Science, Inner Mongolia University, Hohhot 010020, China
2
Jiangsu Provincial Key Laboratory of Biological Therapy for Organ Failure, Nanjing Medical University, Nanjing 211166, China
3
National Center of Technology Innovation for Dairy, Hohhot 010020, China
4
Key Laboratory of Targeted Intervention of Cardiovascular Disease, Collaborative Innovation Center for Cardiovascular Disease Translational Medicine, Nanjing Medical University, Nanjing 211166, China
5
Inner Mongolia Saikexing Institute of Breeding and Reproductive Biotechnology in Domestic Animal, Hohhot 011517, China
*
Authors to whom correspondence should be addressed.
Cells 2026, 15(10), 954; https://doi.org/10.3390/cells15100954 (registering DOI)
Submission received: 8 April 2026 / Revised: 12 May 2026 / Accepted: 20 May 2026 / Published: 21 May 2026
(This article belongs to the Section Stem Cells)
Versions Notes

Abstract

The mammalian early embryo possesses totipotency and can be captured as extended pluripotent stem cells (EPSCs). The first two cell differentiations result in epiblast, primitive endoderm, and trophectoderm, with the trilineage coexisting in a unified uterine microenvironment. Nevertheless, the in vitro counterparts—primed PSCs, trophoblast stem cells (TSCs), and extraembryonic endoderm (XEN) cells—require a distinct culture system. In this study, we successfully derived stable porcine EPSCs from fibroblasts at 35% efficiency, and interestingly observed that these EPSCs differentiated in parallel and gave rise to the transient coexistence (36–84 h) of trilineage cells when cultured in a single system (LCDM: hLIF, CHIR99021, DiM, and MiH). Then, XEN cells gradually predominated and eventually became the sole population in prolonged LCDM culture. However, TSCs and primed PSCs had to be differentiated from EPSCs under their respective culture system. EPSCs can differentiate into PGC-like cells independently of genetic modification and contribute to mouse neurula-stage embryos. Collectively, the trilineage coexistence phenomenon may provide novel insight into an early embryogenesis mechanism and strategy for porcine blastoid construction.
Keywords: porcine; EPSCs; differentiation; trilineage; coexistence porcine; EPSCs; differentiation; trilineage; coexistence

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MDPI and ACS Style

Zhao, L.; Chen, Y.; Guo, X.; Zhou, M.; Guo, T.; Ma, J.; Zhang, M.; Cheng, L.; Yu, J.; Zhang, Y.; et al. The Trilineage Coexistence Observed During the Differentiation of Porcine EPSCs. Cells 2026, 15, 954. https://doi.org/10.3390/cells15100954

AMA Style

Zhao L, Chen Y, Guo X, Zhou M, Guo T, Ma J, Zhang M, Cheng L, Yu J, Zhang Y, et al. The Trilineage Coexistence Observed During the Differentiation of Porcine EPSCs. Cells. 2026; 15(10):954. https://doi.org/10.3390/cells15100954

Chicago/Turabian Style

Zhao, Lihua, Yanglin Chen, Xiyun Guo, Meng Zhou, Tianxu Guo, Junjun Ma, Manling Zhang, Linxin Cheng, Jinbo Yu, Yu Zhang, and et al. 2026. "The Trilineage Coexistence Observed During the Differentiation of Porcine EPSCs" Cells 15, no. 10: 954. https://doi.org/10.3390/cells15100954

APA Style

Zhao, L., Chen, Y., Guo, X., Zhou, M., Guo, T., Ma, J., Zhang, M., Cheng, L., Yu, J., Zhang, Y., Yang, G., Li, R., & Li, X. (2026). The Trilineage Coexistence Observed During the Differentiation of Porcine EPSCs. Cells, 15(10), 954. https://doi.org/10.3390/cells15100954

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