Pluripotent stem cells related to embryonic disc exhibit common self-renewal requirements in diverse livestock species

Kinoshita, Masaki; Kobayashi, Toshihiro; Planells, Benjamin; Klisch, Doris; Spindlow, Daniel; Masaki, Hideki; Bornelöv, Susanne; Stirparo, Giuliano Giuseppe; Matsunari, Hitomi; Uchikura, Ayuko; Lamas-Toranzo, Ismael; Nichols, Jennifer; Nakauchi, Hiromitsu; Nagashima, Hiroshi; Alberio, Ramiro; Smith, Austin

Abstract

Despite four decades of effort, robust propagation of pluripotent stem cells from livestock animals remains challenging. The requirements for self-renewal are unclear and the relationship of cultured stem cells to pluripotent cells resident in the embryo uncertain. Here, we avoided using feeder cells or serum factors to provide a defined culture microenvironment.We show that the combination of activin A, fibroblast growth factor and the Wnt inhibitor XAV939 (AFX) supports establishment and continuous expansion of pluripotent stem cell lines from porcine, ovine and bovine embryos. Germlayer differentiation was evident in teratomas and readily induced in vitro. Global transcriptome analyses highlighted commonality in transcription factor expression across the three species, while global comparison with porcine embryo stages showed proximity to bilaminar disc epiblast. Clonal genetic manipulation and gene targeting were exemplified in porcine stemcells. We further demonstrated that genetically modified AFX stem cells gave rise to cloned porcine foetuses by nuclear transfer. In summary, for major livestockmammals, pluripotent stemcells related to the formative embryonic disc are reliably established using a common and defined signalling environment.

Citation

Kinoshita, M., Kobayashi, T., Planells, B., Klisch, D., Spindlow, D., Masaki, H., Bornelöv, S., Stirparo, G. G., Matsunari, H., Uchikura, A., Lamas-Toranzo, I., Nichols, J., Nakauchi, H., Nagashima, H., Alberio, R., & Smith, A. (2021). Pluripotent stem cells related to embryonic disc exhibit common self-renewal requirements in diverse livestock species. Development, 148(23), Article dev199901. https://doi.org/10.1242/dev.199901

• Journal Article Type: Article
• Acceptance Date: Oct 26, 2021
• Online Publication Date: Dec 7, 2021
• Publication Date: 2021-12
• Deposit Date: May 12, 2022
• Publicly Available Date: May 16, 2022
• Journal: Development (Cambridge)
• Print ISSN: 0950-1991
• Electronic ISSN: 1477-9129
• Publisher: Company of Biologists
• Peer Reviewed: Peer Reviewed
• Volume: 148
• Issue: 23
• Article Number: dev199901
• DOI: https://doi.org/10.1242/dev.199901
• Keywords: Developmental Biology; Molecular Biology
• Public URL: https://nottingham-repository.worktribe.com/output/7017801
• Publisher URL: https://journals.biologists.com/dev/article/148/23/dev199901/273644/Pluripotent-stem-cells-related-to-embryonic-disc