Embryonic stem cells (ESCs) and induced pluripotent stem cells have the potential to differentiate to all cell types of an adult individual and are useful for studying development and for translational research. However, extrapolation of mouse and human ESC knowledge to deriving stable ESC lines of domestic ungulates and large livestock species has been challenging. In contrast to ESCs that are usually established from the blastocyst, mouse expanded potential stem cells (EPSCs) are derived from four-cell and eight-cell embryos. We have recently used the EPSC approach and established stem cells from porcine and human preimplantation embryos. EPSCs are molecularly similar across species and have broader developmental potential to generate embryonic and extraembryonic cell lineages. We further explore the EPSC technology for mammalian species refractory to the standard ESC approaches and report here the successful establishment of bovine EPSCs (bEPSCs) from preimplantation embryos of both wild-type and somatic cell nuclear transfer. bEPSCs express high levels of pluripotency genes, propagate robustly in feeder-free culture, and are genetically stable in long-term culture. bEPSCs have enriched transcriptomic features of early preimplantation embryos and differentiate in vitro to cells of the three somatic germ layers and, in chimeras, contribute to both the embryonic (fetal) and extraembryonic cell lineages. Importantly, precise gene editing is efficiently achieved in bEPSCs, and genetically modified bEPSCs can be used as donors in somatic cell nuclear transfer. bEPSCs therefore hold the potential to substantially advance biotechnology and agriculture.

胚胎干细胞 (ESC) 和诱导多能干细胞具有分化为成年个体所有细胞类型的潜力，可用于研究发育和转化研究。然而，将小鼠和人类 ESC 知识外推到家养有蹄类动物和大型牲畜物种的稳定 ESC 系一直具有挑战性。与通常从囊胚建立的 ESC 不同，小鼠扩增的潜在干细胞 (EPSC) 来自四细胞和八细胞胚胎。我们最近使用 EPSC 方法并从猪和人类植入前胚胎中建立了干细胞。EPSCs 跨物种在分子上相似，并且具有更广泛的发育潜力来产生胚胎和胚胎外细胞谱系。我们进一步探索了适用于标准 ESC 方法难治的哺乳动物物种的 EPSC 技术，并在此报告从野生型和体细胞核移植的植入前胚胎成功建立牛 EPSC（bEPSC）。bEPSCs 表达高水平的多能性基因，在无饲养层培养中繁殖强劲，并且在长期培养中遗传稳定。bEPSCs 丰富了早期植入前胚胎的转录组学特征，并在体外分化为三个体细胞胚层的细胞，并且在嵌合体中，有助于胚胎（胎儿）和胚胎外细胞谱系。重要的是，在 bEPSCs 中有效地实现了精确的基因编辑，并且转基因 bEPSCs 可以用作体细胞核转移的供体。