Isolation and Culture of Embryonic Germ Cells

Abstract

Publisher Summary In mammals, three types of pluripotent stem cells have been identified and isolated into culture; these are embryonic stem (ES) cells, embryonic germ (EG) cells, and embryonal carcinoma (EC) cells. These pluripotent stem cells share two important properties. First, they can be maintained indefinitely in culture as an essentially immortal cell line. Second, they are capable of giving rise to every cell type in the body. These features make such cells potentially important tools for the treatment of human disease because the differentiated derivatives of pluripotent stem cells could be used to replace damaged or diseased cells via transplantation. Pluripotent stem cells and their derivatives will also likely generate important information about embryonic development. Many of the standard techniques used for the culture of ES cells—such as culture medium, feeding regimen, and subculture technique—can be used for the culture of EG cells. The major difference lies in the initial isolation of the cells. EG cell lines have been derived from both mouse and human embryos using the same culture conditions. These observations suggest that some of the factors regulating primordial germ cell (PGC) growth and differentiation have been conserved during evolution and, therefore, that the techniques for isolating EG cells described here may be applicable to other mammalian and nonmammalian species. This chapter outlines the protocols developed for the culture of mouse PGCs and their conversion into EG cells, but the protocols may be applicable to the culture of the same cells from a variety of different species, including birds and humans.