Culture of Postimplantation Mouse Embryos

Abstract

A major disadvantage of working with postimplantation mammalian embryos is their relative inaccessibility to experimentation while they develop within the maternal uterus. Two techniques allow us to get around this problem to a large extent. The first, which is the subject of this chapter, can be used for mouse embryos explanted between 7.5 d of gestation (E7) and 12.5 d of gestation (E12), and involves dissecting embryos from the uterus and culturing them in roller bottles (1). In this way, embryos can be surgically or chemically manipulated or labeled and will develop quite normally in culture for periods of 12–60 h, depending on the stage at explantation (2,3). The second technique, which allows experimentation on more advanced stages, is that of exo utero or open uterus surgery, in which fetuses are suspended in the fluid-filled abdominal cavity of the female mouse while retaining their placental attachment to the uterine wall (4). This procedure is suitable for fetuses at 12.5 d of gestation (E12) and beyond. This chapter will focus on the techniques for culturing E11 mouse embryos with open yolk sacs at limb-bud stages (5) and will also include protocols for culturing earlier stage embryos. We will describe studies in the mouse, but similar manipulations are possible with rat embryos, bearing in mind that they are generally 1 or 2 d behind mouse development; for example, an E11 rat embryo closely resembles a mouse embryo between E9 and E10.