Chapter 21 - Isolation and Maintenance of Primate ES Cells

Abstract

This chapter focuses on techniques for derivation, characterization, maintenance, and differentiation of primate embryonic stem (ES) cells. During the 1970s, multipotent cell lines were isolated from the stem cells of mouse teratocarcinomas. Some of these embryonal carcinoma (EC) cell lines were shown to differentiate in vitro into a variety of cell types, including muscle and nerve cells. When grown in suspension, EC cells tend to aggregate into cell clusters known as embryoid bodies (EBs) in which part of the cells differentiate spontaneously. They also had been shown to differentiate in vivo, forming teratocarcinomas, following their injection into recipient mice. EC cells served for years as models for research on development, establishing all the methodological know-how needed for the isolation and maintenance of ES cells. The ability of nonhuman primate ES cell lines to integrate into all three germ layers during embryonic development using a blastocyst injection model have not been tested. The pluripotency of human postimplantation embryonic cells, between the time of implantation and the gastrulation process, has never been examined previously. Two methods of obtaining such lines are genetic manipulation of existing human ES (hES) cell lines or derivation of hES cell lines from genetically compromised embryos.