Generation of Porcine Blastoids from Embryonic Stem Cells

Stem cell-based blastocyst-like structure models (blastoids) that mimic preimplantation blastocysts can be used to study embryogenesis and key early embryonic developmental events. Large animals may benefit from blastoid models for purposes such as improving and accelerating breeding. We developed a three-dimensional (3D) two-step differentiation strategy to generate porcine blastoids from embryonic stem cells (ESCs). The ESC-derived blastoids exhibit similar morphology, cell lineage composition, and single-cell transcriptome characteristics to porcine blastocysts. These porcine blastoids can survive and expand for more than two weeks in vitro under two different culture conditions. Here, we describe a step-by-step protocol for the generation of porcine blastoids from ESCs.