Self-organization of bovine blatocyst-like structure from three pluripotent stem cells

Here we report the derivation and long-term culture of bovine extraembryonic endoderm cells (bXENs). We showed that bXENs enhance maintain the stemness of bovine ESCs and prevent them from differentiation. In the present of signaling cocktail sustaining bXENs, the growth and progression of epiblasts are also facilitated in the developing pre-implantation embryo. Through 3D assembly of bXENs with bovine ESCs and TSCs, we developed an improved bovine blastocyst like structure (bovine blastoid). The bovine blastoids established in this study represents accessible in vitro models for understanding early embryogeneisis and improving reproductive efficiency in livestock species. Overall design: Bovine blastocyst-like structure generated from three cell lineages (bEPSC, bTSC, bXEN) and day8 bovine blasytocyst were collected for single nuclei isolation followed by snRNA-seq.