5/30/2023 0 Comments Embryonic stem cells![]() Results Identification of a Culture Condition that Maintains Bovine Pluripotency. The availability of bEPSCs, which are robustness in culture, permit efficient genome editing, possess expanded developmental potential, are expected to substantially advance bovine stem cell biology to considerably facilitate selecting for superior animals for farming and to open up opportunities for biotechnology. ![]() In this report, we successfully established and characterized bovine EPSCs (bEPSCs). These cells had the typical mouse- and human-primed ESC properties but morphologically did not form distinct cell colonies, unlike ESCs of other species, and no chimera was generated ( 39). Recently, bovine-primed ESCs were reported ( 39), which represents a major advance. The reported bovine iPSC lines often have leaky expression of the reprogramming genetic factors ( 28– 38). These cells, in general, however, lack the standard pluripotent stem cell criteria: poor derivation efficiencies inability of maintaining pluripotency in long-term culture limited developmental potential in the in vitro and in vivo assays such as chimera generation. Intensive efforts have been made to derive bovine ESCs or to reprogram bovine somatic cells to induced plurpotent stem cells (iPSCs) ( 18– 38). Bovine ESCs would be expected to substantially facilitate genome editing, to accelerate molecular breeding schemes for economic traits, and to provide a platform for investigating the bovine preimplantation development with potential applications in improving cloning. ![]() Mouse EPSCs were reported to self-assemble into blastocyst-like structures ( 17).Ĭattle, or cows, are domesticated bovine farm animals and the most common type of large domesticated ungulates. ![]() They possess robust self-renewal capacity in long-term culture, allow efficient genome editing, and generate both embryonic and extraembryonic cell lineages in vitro and in chimeras in terms of mouse and porcine EPSCs ( 7– 9). EPSCs are molecularly and functionally similar across species. Following this line of reasoning, EPSCs were established by inhibiting molecules and pathways operating in preblastocyst embryos ( 7, 9, 10). We posited that deriving stem cells from earlier preimplantation embryos, for example, mouse four-cell or eight-cell cleavage embryos, might help overcome the substantial species differences reported in the blastocyst-stage embryos ( 11– 16). We and others recently reported establishment of mouse, human, and porcine expanded potential stem cells (EPSCs) ( 7– 10). However, extrapolating the 2i/LIF condition to deriving ESCs of large animals has proven challenging. A significant advance in mouse ESC derivation is to use small molecule inhibitors, specifically the ones targeting Mek1/2 (PD0325901) and GSK3 (Chir99021), in the 2i/LIF naïve ESC culture condition ( 6). bEPSCs therefore hold the potential to substantially advance biotechnology and agriculture.Įmbryonic stem cells (ESCs) of mouse, rat, human, and nonhuman primate species are established from the inner cell mass in the blastocyst ( 1– 5). Importantly, precise gene editing is efficiently achieved in bEPSCs, and genetically modified bEPSCs can be used as donors in somatic cell nuclear transfer. 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. bEPSCs express high levels of pluripotency genes, propagate robustly in feeder-free culture, and are genetically stable in long-term culture. 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. EPSCs are molecularly similar across species and have broader developmental potential to generate embryonic and extraembryonic cell lineages. We have recently used the EPSC approach and established stem cells from porcine and human preimplantation embryos. 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. However, extrapolation of mouse and human ESC knowledge to deriving stable ESC lines of domestic ungulates and large livestock species has been challenging. 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.
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