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Culture of Postimplantation Mouse Embryos

  • Paul Martin
  • David L. Cockroft
Part of the METHODS IN MOLECULAR BIOLOGY™ book series (MIMB, volume 461)

1. Introduction

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...

Keywords

Amniotic Membrane Ectoplacental Cone Iridectomy Scissors Monastral Blue Placental Attachment 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

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.

References

  1. 1.
    New, D. A. T., Coppola, P. T., and Terry, S. (1973) Culture of explanted rat embryos in rotating tubes.J. Reprod. Fert. 35, 135–138.CrossRefGoogle Scholar
  2. 2.
    New, D. A. T. (1978) Whole-embryo culture and the study of mammalian embryos during organogenesis.Biol. Rev. 53, 81–122.CrossRefPubMedGoogle Scholar
  3. 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. 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
  5. 5.
    Cockroft, D. L. (1973) Development in culture of rat foetuses explanted at 12.5 and 13.5 days of gestation.J. Embryol. Exp. Morph. 29, 473–483.PubMedGoogle Scholar
  6. 6.
    Sadler, T. W. and New, D. A. T. (1981) Culture of mouse embryos during neurula-tion.J. Embryol. Exp. Morph. 66, 109–116.PubMedGoogle Scholar
  7. 7.
    Steele, C. E. and New, D. A. T. (1974) Serum variants causing the formation of double hearts and other abnormalities in explanted rat embryos.J. Embryol. Exp. Morph. 31, 707–719.PubMedGoogle Scholar
  8. 8.
    New, D. A. T., Coppola, P. T., and Cockroft, D. L. (1976) Improved development of head-fold rat embryos in culture resulting from low oxygen and modifications of the culture serum.J. Reprod. Fert. 48, 219–222.CrossRefGoogle Scholar
  9. 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. 10.
    Kaufman, M. H. (1992)The Atlas of Mouse Development. Academic Press, London.Google Scholar
  11. 11.
    Theiler, K. (1989)The House Mouse. Atlas of Embryonic Development. Springer-Verlag, New York.Google Scholar
  12. 12.
    Rugh, R. (1990)The Mouse. Its Reproduction and Development. Oxford University Press, Oxford.Google Scholar
  13. 13.
    Brown, N. A. and Fabro, S. (1981) Quantitation of rat embryonic development in vitro: a morphological scoring system.Teratology 24, 65–78.CrossRefPubMedGoogle Scholar
  14. 14.
    Downs, K. and Davies, T. (1993) Staging of gastrulating mouse embryos by morphological landmarks in the dissecting microscope.Development 118, 1255– 1266.PubMedGoogle Scholar
  15. 15.
    Wanek, N., Muneoka, K., Holler-Dinsmore, G., Burton, R., and Bryant, S. V. (1989) A staging system for mouse limb development.J. Exp. Zool. 249, 41–49.CrossRefPubMedGoogle Scholar
  16. 16.
    Martin, P. (1990) Tissue patterning in the developing mouse limb.Int. J. Dev. Biol. 34, 323–336.PubMedGoogle Scholar
  17. 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
  18. 18.
    Martin, P. and Swanson, G. (1993) Descriptive and experimental analysis of the epithelial remodellings that control semicircular canal formation in the developing mouse inner ear.Dev. Biol. 159, 549–558.CrossRefPubMedGoogle Scholar
  19. 19.
    Martin, P. Dickson, M. C., Millan, F. A., and Akhurst, R. J. (1993) Rapid induction and clearance of TGFβ1 is an early response to wounding in the mouse embryo.Dev. Genet. 14, 225–238.CrossRefPubMedGoogle Scholar
  20. 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
  21. 21.
    McCluskey, J. and Martin, P. (1995) Analysis of the tissue movements of embryonic wound healing—DiI studies in the wounded E11.5 mouse embryo.Dev. Biol. 170, 102–114.CrossRefPubMedGoogle Scholar
  22. 22.
    Cockroft, D. L. (1988) Changes with gestational age in the nutritional requirements of postimplantation rat embryos in culture.Teratology 38, 281–290.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science + Business Media, LLC 2008

Authors and Affiliations

  • Paul Martin
    • 1
  • David L. Cockroft
    • 2
  1. 1.Departments of Physiology and Biochemistry, School of Medical SciencesUniversity of BristolBristolUK
  2. 2.Formerly Imperial Cancer Research FundOxfordUK

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