Skip to main content

Isolation, Culture, and Characterisation of Mouse Embryonic Oesophagus and Intestine

  • Protocol
  • First Online:
Book cover Mouse Cell Culture

Part of the book series: Methods in Molecular Biology ((MIMB,volume 633))

  • 9531 Accesses

Abstract

The gastrointestinal tract of vertebrates is lined by epithelium that develops from the endodermal germ layer. The oesophagus and intestine form part of the gastrointestinal tract and studying the normal development of both tissues is difficult due to lack of suitable in vitro model systems. One of the criteria for a reliable culture model includes the ability to carry out real-time observations in vitro. The method we describe here is based on the isolation of embryonic oesophagus and intestine from 11.5-day-old embryos and culture on fibronectin-coated coverslips in basal Eagle’s medium and 20% fetal bovine serum. This model permits real-time observations of both tissues and over a few days in culture, markers of differentiation appear. This culture system appears to recapitulate normal oesophagus and intestine development.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Slack, J. M. W. (2006) Development of endodermal organs. In Essential develop- mental biology, 2nd ed. Blackwell Scientific, Oxford, pp. 249–263.

    Google Scholar 

  2. Yu, W.-Y., Slack, J. M. W., and Tosh, D. (2005) Conversion of columnar to stratified squamous epithelium in the developing mouse oesophagus. Dev. Biol. 284, 157–170.

    Article  CAS  PubMed  Google Scholar 

  3. Duan, H., Gao, F., Li, S., and Nagata, T. (1993) Postnatal development and aging of esophageal epithelium in mouse: a light and electron microscopic radioautographic study. Cell. Mol. Biol. 39, 309–316.

    CAS  PubMed  Google Scholar 

  4. Mathan, M., Moxey, P. C., and Trier, J. S. (1976) Morphogenesis of fetal rat duodenal villi. Am. J. Anat. 146, 73–92.

    Article  CAS  PubMed  Google Scholar 

  5. Marshman, E., Booth, C., and Potten, C. S. (2002) The intestinal epithelial stem cell. BioEssays 24, 91–98.

    Article  PubMed  Google Scholar 

  6. Hall, P. A., Coates, P. J., Ansari, B., and Hopwood, D. (1994) Regulation of cell number in the mammalian gastrointestinal tract: the importance of apoptosis. J. Cell Sci. 107, 3569–3577.

    CAS  PubMed  Google Scholar 

  7. Strater, J., Koretz, K., Gunthert, A. R., and Moller, P. (1995) In situ detection of enterocytic apoptosis in normal colonic mucosa and in familial adenomatous polyposis. Gut 37, 819–825.

    Article  CAS  PubMed  Google Scholar 

  8. Tait, I. S., Evans, G. S., Kedinger, M., Flint, N., Potten, C. S., and Campbell, F. C. (1994) Progressive morphogenesis in vivo after transplantation of cultured small bowel epithelium. Cell. Transplant. 3, 33–40.

    CAS  PubMed  Google Scholar 

  9. Hearn, C. J., Young, H. M., Ciampoli, D., Lomax, A. E., and Newgreen, D. (1999) Catenary cultures of embryonic gastrointestinal tract support organ morphogenesis, motility, neural crest cell migration, and cell differentiation. Dev. Dyn. 214, 239–247.

    Article  CAS  PubMed  Google Scholar 

  10. Abud, H. E., Lock, P., and Heath, J. K. (2004) Efficient gene transfer into the epithelial cell layer of embryonic mouse intestine using low-voltage electroporation. Gastroenterology 126, 1779–1787.

    Article  CAS  PubMed  Google Scholar 

  11. Abud, H. E., Watson, N., and Heath, J. K. (2005) Growth of intestinal epithelium in organ culture is dependent on EGF signaling. Exp. Cell Res. 303, 252–262.

    Article  CAS  PubMed  Google Scholar 

  12. Percival, A. C., and Slack, J. M. W. (1999) Analysis of pancreatic development using a cell lineage label. Exp. Cell Res. 247, 123–132.

    Article  CAS  PubMed  Google Scholar 

  13. Shen, C. -N., Slack, J. M. W., and Tosh, D. (2000) Molecular basis of transdifferentiation of pancreas to liver. Nat. Cell Biol. 2, 879–887.

    Article  CAS  PubMed  Google Scholar 

  14. Shen, C.-N., Seckl, J. R., Slack, J. M. W., and Tosh, D. (2003) Glucocorticoids suppress beta-cell development and induce hepatic metaplasia in embryonic pancreas. Biochem. J. 375, 41–50.

    Article  CAS  PubMed  Google Scholar 

  15. Quinlan, J. M., Yu, W.-Y., Hornsey, M. A., Tosh, D., and Slack, J. M. W. (2006) In vitro culture of mouse intestinal epithelium: cell differentiation and introduction of reporter genes. BMC Dev. Biol. 6, 24.

    Article  PubMed  Google Scholar 

  16. Real, F. X., Xu, M., Vila, M. R., and de Bolos, C. (1992) Intestinal brush-border associated enzymes: co-ordinated expression in colorectal cancer. Int. J. Cancer 51, 173–181.

    Article  CAS  PubMed  Google Scholar 

  17. Hocker, M., and Wiedenmann, B. (1998) Molecular mechanisms of enteroendocrine differentiation. Ann. NY Acad. Sci. 859, 160–174.

    Article  CAS  PubMed  Google Scholar 

  18. Roth, K. A., and Gordon, J. L. (1990) Spatial differentiation of the intestinal epithelium: analysis of enteroendocrine cells containing immunoreactive serotonin, secretin, and substance P in normal and transgenic mice. Proc. Natl. Acad. Sci. USA 87, 6408–6412.

    Article  CAS  PubMed  Google Scholar 

  19. Porter, E. M., Bevins, C. L., Ghosh, D., and Ganz, T. (2002) The multifaceted Paneth cell. Cell Mol. Life Sci. 59, 156–170.

    Article  CAS  PubMed  Google Scholar 

  20. Silberg, D. G., Swain, G. P., Suh, E. R., and Traber, P. G. (2000) Cdx1 and Cdx2 expression during intestinal development. Gastroenterology 119, 961–971.

    Article  CAS  PubMed  Google Scholar 

  21. Owens, D. W. and Lane, E. B. (2003) The quest for the function of simple epithelial keratins. BioEssays 25, 748–758.

    Article  CAS  PubMed  Google Scholar 

  22. Boukhalfa, G., Desmouliere, A., Rondeau, E., Gabbiani, G., and Sraer, J. D. (1996) Relationship between alpha-smooth muscle actin expression and fibrotic changes in human kidney. Exp. Nephrol. 4, 241–247.

    CAS  PubMed  Google Scholar 

  23. Reid, P., Owen, D. A., Magee, F., and Park, C. M. (1990) Histochemical studies of intestinal epithelial goblet cell glycoproteins during the development of the human foetus. Histochem. J. 22, 81–86.

    Article  CAS  PubMed  Google Scholar 

  24. Hawkins, K. L., Lloyd, R. V., and Toy, K. A. (1989) Immunohistochemical localization of chromogranin A in normal tissues from laboratory animals. Vet. Pathol. 26, 488–498.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

The authors are grateful to Cancer Research UK, for funding this work.

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2010 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Quinlan, J.M., Yu, WY., Tosh, D. (2010). Isolation, Culture, and Characterisation of Mouse Embryonic Oesophagus and Intestine. In: Ward, A., Tosh, D. (eds) Mouse Cell Culture. Methods in Molecular Biology, vol 633. Humana Press. https://doi.org/10.1007/978-1-59745-019-5_6

Download citation

  • DOI: https://doi.org/10.1007/978-1-59745-019-5_6

  • Published:

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-772-3

  • Online ISBN: 978-1-59745-019-5

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics