Culture of Postimplantation Mouse Embryos
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...
KeywordsAmniotic Membrane Ectoplacental Cone Iridectomy Scissors Monastral Blue Placental Attachment
We are particularly indebted to Catherine Haddon for the lucid diagrams illustrating the dissections. We also thank The Medical Research Council and the Wellcome Trust (P. M.), and the Imperial Cancer Research Fund (D. L. C.) for financial support.
- 3.Cockroft, D. L. (1990) Dissection and culture of postimplantation embryos, inPostimplantation Mammalian Embryos—A Practical Approach(Copp, A. J. and Cockroft, D. L., eds.), IRL, Oxford, pp. 15–40.Google Scholar
- 4.Muneoka, K., Wanek, N., Trevino, C., and Bryant, S. V. (1990) Exo utero surgery, inPostimplantation Mammalian Embryos—A Practical Approach(Copp, A. J. and Cockroft, D. L., eds.), IRL, Oxford, pp. 41–59.Google Scholar
- 9.Freeman, S. J., Coakley, M. E., and Brown, N. A. (1987) Post-implantation embryo culture for studies of teratogenesis, inBiochemical Toxicology—A Practical Approach(Snell, K. and Mullock, B., eds.), IRL, Oxford, UK, pp. 83–107.Google Scholar
- 10.Kaufman, M. H. (1992)The Atlas of Mouse Development. Academic Press, London.Google Scholar
- 11.Theiler, K. (1989)The House Mouse. Atlas of Embryonic Development. Springer-Verlag, New York.Google Scholar
- 12.Rugh, R. (1990)The Mouse. Its Reproduction and Development. Oxford University Press, Oxford.Google Scholar
- 17.Karnovsky, M. J. (1965) A formaldehyde-gluteraldehyde fixative of high osmolar-ity for use in electron microscopy.J. Cell Biol. 27, 137,138.Google Scholar
- 20.Hopkinson-Woolley, J., Hughes, D. A., Gordon, S., and Martin, P. (1994) Macro-phage recruitment during limb development and wound healing in the embryonic and foetal mouse.J. Cell Sci. 170, 1159–1167.Google Scholar