Avian embryonic stem cells.

Blastodermal cells derived from the area pellucida of a stage X (EG&K) embryo have the potential to contribute to the somatic tissues and the germ line when reintroduced into a stage X (EG&K) recipient embryo. This chapter describes a method to culture chicken embryonic stem (cES) cells derived from blastodermal cells. Within the first week of culture, the cells change their morphology; they become smaller with a large nucleus and a prominent nucleolus. The cES cells remain chromosomally normal and can be cultured for extended periods. They can be modified genetically using standard electroporation procedures and, after injection into a recipient embryo, can contribute to all somatic tissues. Using a surrogate shell culture system, the injected embryos can be manipulated and visualized easily throughout incubation. We have generated high-grade chimeras by compromising the recipient embryos and maintaining the ES cells in stage X (EG&K) recipients for a few days at 15 degrees before incubating them at 37.5 degrees. The cES system provides a novel experimental paradigm for the investigation of developmental and physiological mechanisms in the chicken.