Development of Adipocytes from Differentiated ES Cells

Abstract

This chapter describes an in vitro system that allows access to the development of adipocytes from pluripotent mouse ES cells. Because there are no specific markers that can be used to identify adipoblasts and preadipocytes in vivo , the white adipose tissue can be identified only when it contains adipocytes (i.e., terminally differentiated cells) and cannot be detected macroscopically during embryogenesis. Therefore, very little is known about the ontogeny of adipose tissue. Mainly, the in vitro system used to study adipogenesis is immortal preadipocyte cell lines, which represent already determined cells. The use of in vitro differentiation of ES cells to analyze the effects of mutations on adipogenesis has begun recently and master genes that commit progression from pluripotent stem cells to the adipogenic lineage have not yet been identified. The chapter also describes a protocol for culture conditions in which ES cells undergo adipocyte differentiation at a high rate and in a highly reproducible fashion. This culture system should provide a powerful means for studying the first step of adipose cell development and a route to identify regulatory genes involved in adipogenesis.