Regulation of early development can be directly interrogated in the embryo or can be studied in cultured cells isolated in the laboratory. New understanding of the developmental stages and the signalling requirements of the pig embryo have enabled the development of improved protocols for the derivation of pluripotent cells from early epiblasts. Here, we provide a detailed step-by-step description of the critical parameters required for isolation and establishment of these cells and how they can be used to study their developmental properties.