ESTABLISHMENT, MAINTENANCE, AND TRANSFECTION OF TOTIPOTENT EMBRYONIC STEM CELLS FROM THE EMBRYOS OF DOMESTIC ANIMALS

摘要

A method is described for establishing embryonic stem (ES) cell lines of domestic animals, such as cattle, sheep, goats, rabbits, and mink. Expanded blastocysts of the domestic animal are treated to remove the zonae pellucidae and then seeded onto a layer of feeder cells in a culture dish containing a suitable medium. The medium comprises plasma that is low in mitogens instead of serum that has undergone the clotting cascade. Addition of leukemia inhibitory factor (LIF), alpha-fetoprotein, and/or hyaluronic acid to the medium also improves the efficiency of establishing such ES cell line. Mink ES cell lines are prepared by pretreating blastocysts with prolactin to reactivate the blastocysts from embryonic diapause. Transfection of such ES cell lines and a composition for use in establishing embryonic stem cell lines are also disclosed.