While research into deriving and manipulating pluripotent stem cells (PSCs) from humans and mice has proceeded at a rapid pace, the robust propagation of PSCs from livestock has lagged behind. Now, Hiroshi Nagashima, Ramiro Alberio and Austin Smith present a protocol for the establishment and continuous expansion of PSC lines from porcine, ovine and bovine embryos in defined culture conditions. Transcriptome analysis identifies a common transcription factor expression profile between the species. Further, when compared with porcine embryo stages, the global transcriptome of porcine PSCs shows proximity to that of the bilaminar disc epiblast prior to gastrulation, leading the authors to refer to these in vitro-derived cells collectively as embryonic disc stem cells (EDSCs). The authors show that EDSCs are competent to differentiate into all three germ layers and, promisingly for a potential role in in vitro meat production, they were able to push the bovine EDSCs to form myotubes. Finally, they demonstrate that porcine EDSCs are amenable to genetic manipulation using CRISPR/Cas9 gene editing, and can be used as donors for nuclear transfer. The ability to derive and maintain PSCs from livestock paves the way for new research in comparative mammalian embryology, sustainable meat production, enhanced genetic selection of livestock and xenotransplantation.