Open AccessPluripotent stem cells related to embryonic disc exhibit common self-renewal requirements in diverse livestock species
Author(s) -
Masaki Kinoshita,
Toshihiro Kobayashi,
Benjamín Planells,
Doris Klisch,
Daniel Spindlow,
Masaki Hara,
Susanne Bornelöv,
Giuliano Giuseppe Stirparo,
Hitomi Matsunari,
Ayuko Uchikura,
Ismael Lamas-Toranzo,
Jennifer Nichols,
Hiromitsu Nakauchi,
Hiroshi Nagashima,
Ramiro Alberio,
Austin Smith
Publication year - 2021
Publication title -
development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.754
H-Index - 325
eISSN - 1477-9129
pISSN - 0950-1991
DOI - 10.1242/dev.199901
Subject(s) - biology , induced pluripotent stem cell , embryonic stem cell , stem cell , epiblast , microbiology and biotechnology , germ layer , embryoid body , wnt signaling pathway , transcriptome , kosr , cellular differentiation , genetics , immunology , gene expression , gene , gastrulation , signal transduction
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. Germ layer 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 stem cells. We further demonstrated that genetically modified AFX stem cells gave rise to cloned porcine foetuses by nuclear transfer. In summary, for major livestock mammals, pluripotent stem cells related to the formative embryonic disc are reliably established using a common and defined signalling environment. This article has an associated ‘The people behind the papers’ interview.