Isolation and Culture of Embryonic Pancreas and Liver

  • Zoë D. Burke
  • Wan-Chun Li
  • Jonathan M.W. Slack
  • David Tosh
Part of the Methods in Molecular Biology book series (MIMB, volume 633)


Culturing embryonic tissue in an in vitro setting offers the unique ability to manipulate the external medium and therefore to investigate the pathways involved in regulating normal organogenesis as well as providing models for developmental disorders. Here we describe a system for the in vitro culture of the dorsal pancreatic buds and liver buds from mouse embryos. The tissues are dissected from day 9.0 or 11.5 mouse embryos. The tissues are placed on fibronectin-coated coverslips in serum-containing medium and allowed to attach. Over the next few days, the buds grow as flattened structures which are thin enough to allow the use of wholemount immunostaining methods.

Key words

Pancreas liver embryo culture 



The authors are grateful to the Wellcome Trust for funding this work.


  1. 1.
    Jaskoll, T., Leo, T., Witcher, D., Ormestad, M., Astorga, J., Bringas, P., Carlsson, P., and Melnick, M. (2004) Sonic hedgehog signalling plays an essential role during embryonic salivary gland epithelial branching morphogenesis. Dev. Dyn. 229, 722–732.CrossRefPubMedGoogle Scholar
  2. 2.
    Esni, F., Ghosh, B., Biankin, A. V., Lin, J. W., Albert, M. A., Yu, X. et al. (2004) Notch inhibits Ptf1 function and acinar cell differentiation in developing mouse and zebrafish pancreas. Development 131, 4213–4224.CrossRefPubMedGoogle Scholar
  3. 3.
    Hussain, S. Z., Sneddon, T., Tan, X., Micsenyi, A., Michalopoulos, G. K., and Monga, S. P. S. (2004) Wnt impacts growth and differentiation in ex vivo liver development. Exp. Cell Res. 292(1), 157–169.CrossRefPubMedGoogle Scholar
  4. 4.
    Weaver, M., Yingling, J. M., Dunn, N. R., Bellusci, S., and Hogan, B. L. (1999) Bmp signaling regulates proximal-distal differentiation of endoderm in mouse lung development. Development 126(18), 4005–4015.PubMedGoogle Scholar
  5. 5.
    Davies, J. A., Ladomery, M., Hohenstein, P., Michael, L., Shafe, A., Spraggon, L., and Hastie, N. (2004) Development of an siRNA-based method for repressing specific genes in renal cultures and its use to show that the WT1 tumor suppressor is required for nephron differentiation. Human Mol. Gen. 13(2), 235–246.CrossRefGoogle Scholar
  6. 6.
    Jung, J., Zheng, M., Goldfarb, M., and Zaret, K. S. (1999) Initiation of mammalian liver development from endoderm by fibroblast growth factors. Science 284, 1998–2003.CrossRefPubMedGoogle Scholar
  7. 7.
    Rossi, J. M., Dunn, N. R., Hogan, B. L. M., and Zaret, K. S. (2001) Distinct mesodermal signals, including BMPs from the septum transversum mesenchyme, are required in combination for hepatogenesis from the endoderm. Genes Dev. 15, 1998–2009.CrossRefPubMedGoogle Scholar
  8. 8.
    Duncan, S. (2003) Mechanisms controlling early development of the liver. Mech. Dev. 120, 19–33.CrossRefPubMedGoogle Scholar
  9. 9.
    Lemaigre, F. (2003) Development of the biliary tract. Mech. Dev. 120, 81–87.CrossRefPubMedGoogle Scholar
  10. 10.
    Jensen, J. (2004) Gene regulatory factors in pancreatic development. Dev. Dyn. 229, 176–200.CrossRefPubMedGoogle Scholar
  11. 11.
    Murtaugh, L. C., and Melton, D. A. (2003) Genes, signals and lineages in pancreas development. Annu. Rev. Cell Dev. Biol. 19, 71–89.CrossRefPubMedGoogle Scholar
  12. 12.
    Yoshitomi, H., and Zaret, K. S. (2004) Endothelial cell interactions initiate dorsal pancreas development by selectively inducing the transcription factor Ptf1a. Development 131, 807–817.CrossRefPubMedGoogle Scholar
  13. 13.
    Kim, S. K., Hebrok, M., and Melton, D. A. (1997) Notochord to endoderm signalling is required for pancreas development. Development 124, 4243–4252.PubMedGoogle Scholar
  14. 14.
    Percival, A. C., and Slack, J. M. W. (1999) Analysis of pancreatic development using a cell lineage label. Exp. Cell Res. 247, 123–132.CrossRefPubMedGoogle Scholar
  15. 15.
    Kurash, J. K., Shen, C.-N., and Tosh, D. (2004) Induction and regulation of acute phase proteins in transdifferentiated hepatocytes. Exp. Cell Res. 292, 342–358.CrossRefPubMedGoogle Scholar
  16. 16.
    Shen, C.-N., Marguerie, A., Chien, C.-Y., Dickson, C., Slack, J. M. W., and Tosh, D. (2007) All- trans retinoic acid suppresses exocrine differentiation and branching morphogenesis in the embryonic pancreas. Differentiation 75, 62–74.CrossRefPubMedGoogle Scholar
  17. 17.
    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.CrossRefPubMedGoogle Scholar
  18. 18.
    Thowfeequ, S., Ralphs, K. L., Yu, W.-Y., Slack, J. M. W., and Tosh, D. (2007) Betacellulin inhibits amylase and glucagon production and promotes beta-cell production in mouse embryonic pancreas. Diabetologia 50, 1688–16977.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Zoë D. Burke
    • 1
  • Wan-Chun Li
    • 1
  • Jonathan M.W. Slack
    • 1
    • 2
  • David Tosh
    • 1
  1. 1.Department of Biology and BiochemistryCentre for Regenerative Medicine, University of BathBathUK
  2. 2.Stem Cell Institute, University of MinnesotaMinneapolisUSA

Personalised recommendations