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Conversion between porcine naïve-like and primed ESCs and specific pluripotency marker identification

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Abstract

Researchers currently lack standardized porcine-specific markers that would aid in distinguishing the naïve and primed states of porcine embryonic stem cells (ESCs). Here, we converted naïve-like porcine ESCs (nESCs, established in our lab) into primed-state cells, and we proposed a set of molecular criteria for evaluating the naïve porcine ESCs by comparing the two cell states. The reverse-primed porcine ESCs (rpESCs) are phenotypically stable and karyotypically intact. Alkaline phosphatase positivity and the ability to form embryonic bodies suggest that rpESCs still retain the capacity for self-renewal. Lineage-associated genes, such as Cdx2, Sox17, Eomes, Foxa, Fgf5, and Pitx2, exhibited significant expression in rpESCs. Nonetheless, LIF/3i-grown porcine ESCs treated with the small molecular weight inhibitors CHIR99021, PD0325901, and SB431542 expressed the greatest number of pluripotency marker genes, including Oct4, Sox2, Nog, Dppa5, Nr0b1, and Klf4, and at higher levels than were observed in rpESCs. Despite their general trend toward higher expression of critical pluripotency factors, the nESCs showed downregulation of Tbx3, Nanog, and c-Myc, which are considered typical naïve factors in other species. Entry of the nESCs into the developmentally primed state was also associated with a marked reduction in Lin28 expression. These findings extend the knowledge of porcine pluripotency markers and provide a backdrop for future analysis of naïve porcine pluripotency.

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Funding

We received grants from the Collaborative Innovation Center for Cardiovascular Disease Translational Medicine, Jiangsu Province, China. This work was supported by the National Natural Sciences Foundation of China (31371487).

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Authors and Affiliations

  1. Jiangsu Key Laboratory of Xenotransplantation, Nanjing Medical University, Nanjing, Jiangsu, China

    Qiaoyu Chen, Hong Zhang, Haibin Jiang, Manling Zhang, Junzheng Wang, Lihua Zhao, Chenyu Wang, Manling Liu & Rongfeng Li

  2. State Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, Jiangsu, China

    Qiaoyu Chen, Hong Zhang, Haibin Jiang, Manling Zhang, Junzheng Wang, Lihua Zhao, Chenyu Wang, Manling Liu & Rongfeng Li

  3. Key Laboratory of Targeted Intervention of Cardiovascular Disease, Collaborative Innovation Center for Cardiovascular Disease Translational Medicine, Nanjing Medical University, Nanjing, Jiangsu, China

    Manling Zhang, Lihua Zhao & Rongfeng Li

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  1. Qiaoyu Chen
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  2. Hong Zhang
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  3. Haibin Jiang
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  9. Rongfeng Li
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Correspondence to Rongfeng Li.

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Chen, Q., Zhang, H., Jiang, H. et al. Conversion between porcine naïve-like and primed ESCs and specific pluripotency marker identification. In Vitro Cell.Dev.Biol.-Animal 56, 412–423 (2020). https://doi.org/10.1007/s11626-020-00448-3

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