Gene

Cell biology

Demography

Induced pluripotent stem cell

Biology

Blastocyst

Stem cell

Inner cell mass

Genetics

Transcriptome

Somatic cell nuclear transfer

Embryo

Genome

Embryonic stem cell

Embryogenesis

Population

Sociology

Bovine genome

Gene expression

Wnt signaling pathway

Proceedings of the National Academy of Sciences

Embryonic stem cells (ESCs) are derived from the inner cell mass of preimplantation blastocysts. From agricultural and biomedical perspectives, the derivation of stable ESCs from domestic ungulates is important for genomic testing and selection, genome engineering, and modeling human diseases. Cattle are one of the most important domestic ungulates that are commonly used for food and bioreactors. To date, however, it remains a challenge to produce stable pluripotent bovine ESC lines. Employing a culture system containing fibroblast growth factor 2 and an inhibitor of the canonical Wnt-signaling pathway, we derived pluripotent bovine ESCs (bESCs) with stable morphology, transcriptome, karyotype, population-doubling time, pluripotency marker gene expression, and epigenetic features. Under this condition bESC lines were efficiently derived (100% in optimal conditions), were established quickly (3-4 wk), and were simple to propagate (by trypsin treatment). When used as donors for nuclear transfer, bESCs produced normal blastocyst rates, thereby opening the possibility for genomic selection, genome editing, and production of cattle with high genetic value.

Efficient derivation of stable primed pluripotent embryonic stem cells from bovine blastocysts