Pancreas Development Ex Vivo: Culturing Embryonic Pancreas Explants on Permeable Culture Inserts, with Fibronectin-Coated Glass Microwells, or Embedded in Three-Dimensional Matrigel™

  • Hung Ping Shih
  • Maike SanderEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1210)


Pancreas development is a complex and dynamic process orchestrated by cellular and molecular events, including morphogenesis and cell differentiation. As a result of recent explorations into possible cell-therapy-based treatments for diabetes, researchers have made significant progress in deciphering the developmental program of pancreas formation. In vitro pancreas organ culture systems provide a valuable tool for exploring the mechanisms of gene regulation, cellular behaviors, and cell differentiation. In this chapter, we review three common techniques for culturing embryonic pancreas explants. Each technique is suitable for different applications. Specifically, culturing embryonic pancreas on culture inserts provides an excellent platform to test the effects of chemical compounds. Conversely, when the embryonic pancreas is cultured in fibronectin-coated glass microwells, the system provides unique culture conditions to monitor organ growth and cellular dynamic events. Lastly, when the embryonic pancreas is embedded in Matrigel, organogenesis can be studied in a three-dimensional environment instead of limiting the analysis to one plane.

Key words

Pancreas Development Explants Differentiation Morphogenesis 



This work was supported by the NIH/NIDDK awards to M.S. (R01-DK078803 and R01-DK68471), and JDRF postdoctoral fellowships H.P.S. (3-2009-161).


  1. 1.
    Villasenor A, Chong DC, Henkemeyer M, Cleaver O (2010) Epithelial dynamics of pancreatic branching morphogenesis. Development 137:4295–4305PubMedCentralPubMedCrossRefGoogle Scholar
  2. 2.
    Kesavan G, Sand FW, Greiner TU, Johansson JK, Kobberup S et al (2009) Cdc42-mediated tubulogenesis controls cell specification. Cell 139:791–801PubMedCrossRefGoogle Scholar
  3. 3.
    Shih HP, Wang A, Sander M (2013) Pancreas organogenesis: from lineage determination to morphogenesis. Annu Rev Cell Dev Biol 29: 81–105PubMedCrossRefGoogle Scholar
  4. 4.
    Pan FC, Wright C (2011) Pancreas organogenesis: from bud to plexus to gland. Dev Dyn 240:530–565PubMedCrossRefGoogle Scholar
  5. 5.
    Esni F, Miyamoto Y, Leach SD, Ghosh B (2005) Primary explant cultures of adult and embryonic pancreas. Methods Mol Med 103:259–271PubMedGoogle Scholar
  6. 6.
    Petzold KM, Spagnoli FM (2012) A system for ex vivo culturing of embryonic pancreas. J Vis Exp 66:e3979PubMedGoogle Scholar
  7. 7.
    Burke ZD, Li WC, Slack JM, Tosh D (2010) Isolation and culture of embryonic pancreas and liver. Methods Mol Biol 633:91–99PubMedCrossRefGoogle Scholar
  8. 8.
    Packard A, Georgas K, Michos O, Riccio P, Cebrian C et al (2013) Luminal mitosis drives epithelial cell dispersal within the branching ureteric bud. Dev Cell 27:319–330PubMedCrossRefGoogle Scholar
  9. 9.
    Greggio C, De Franceschi F, Figueiredo-Larsen M, Gobaa S, Ranga A et al (2013) Artificial three-dimensional niches deconstruct pancreas development in vitro. Development 140:4452–4462PubMedCrossRefGoogle Scholar
  10. 10.
    Jin L, Feng T, Shih HP, Zerda R, Luo A et al (2013) Colony-forming cells in the adult mouse pancreas are expandable in Matrigel and form endocrine/acinar colonies in laminin hydrogel. Proc Natl Acad Sci U S A 110: 3907–3912PubMedCentralPubMedCrossRefGoogle Scholar
  11. 11.
    Puri S, Hebrok M (2007) Dynamics of embryonic pancreas development using real-time imaging. Dev Biol 306:82–93PubMedCentralPubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  1. 1.Department of Pediatrics and Cellular and Molecular Medicine, Pediatric Diabetes Research Center, UCSD Stem Cell ProgramUniversity of California San DiegoLa JollaUSA

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