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Inhibition of TGF-β pathway improved the pluripotency of porcine pluripotent stem cells

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Abstract

Porcine pluripotent stem cells had been derived from different culture systems. PeNK6 is a porcine pluripotent stem cell line that we established from an E5.5 embryo in a defined culture system. Signaling pathways related with pluripotency had been assessed in this cell line, and TGF-β signaling pathway–related genes were found upregulated significantly. In this study, we elucidated the role of the TGF-β signaling pathway in PeNK6 through adding small molecule inhibitors, SB431542 (KOSB) or A83-01 (KOA), into the original culture medium (KO) and analyzing the expression and activity of key factors involved in the TGF-β signaling pathway. In KOSB/KOA medium, the morphology of PeNK6 became compact and the nuclear-to-cytoplasm ratio was increased. The expression of the core transcription factor SOX2 was significantly upregulated compared with cell lines in the control KO medium, and the differentiation potential became balanced among three germ layers rather than bias to neuroectoderm/endoderm as the original PeNK6 did. The results indicated that inhibition of TGF-β has positive effects on the porcine pluripotency. Based on these results, we established a pluripotent cell line (PeWKSB) from E5.5 blastocyst by employing TGF-β inhibitors, and the cell line showed improved pluripotency.

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Data availability

All data supporting the findings of this study are available in the manuscript, as well as from the corresponding author upon reasonable request.

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Acknowledgements

We thank Renyue Wei, Jiawei Lv, and Jiaqiang Wang for their technical assistance. This work was supported by the National Key Research and Development program of China-Stem cell and Translational Research (2016YFA0100200).

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  1. Fang Gao and Shuang Wu contributed equally to this work.

Authors and Affiliations

  1. College of Life Science, Northeast Forestry University, Harbin, 150040, People’s Republic of China

    Fang Gao & Tiezhu An

  2. Laboratory of Embryo Biotechnology, College of Life Science, Northeast Agricultural University, Harbin, 150030, People’s Republic of China

    Fang Gao, Shuang Wu, Yan Li, Yuan Fang, Minli Liu, Jiawei Du & Qingran Kong

  3. Wenzhou Medical University, Wenzhou, 325035, People’s Republic of China

    Qingran Kong

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  1. Fang Gao
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Contributions

Fang Gao and Shuang Wu conceived the study, designed and carried out the experiments, and drafted the manuscript; Yan Li, Yuan Fang, Minli Liu, Jiawei Du, and Qingran Kong carried out the experiments and participated in data analysis; Tiezhu An supervised the study. All authors have read, discussed, and approved the final manuscript.

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Correspondence to Tiezhu An.

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All animal experiments were approved by the Animal Care and Use Committee of Northeast Forestry University and the Animal Care and Use Committee of Northeast Agricultural University and were performed according to the guidelines for the welfare and use of animals in stem cell research.

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The authors declare no competing interests.

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Gao, F., Wu, S., Li, Y. et al. Inhibition of TGF-β pathway improved the pluripotency of porcine pluripotent stem cells. In Vitro Cell.Dev.Biol.-Animal 59, 142–152 (2023). https://doi.org/10.1007/s11626-023-00752-8

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