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Isolation, culture and genetic manipulation of mouse pancreatic ductal cells


The most common subtype of pancreatic cancer is pancreatic ductal adenocarcinoma (PDAC). PDAC resembles duct cells morphologically and, to some extent, at a molecular level. Recently, genetic-lineage labeling has become popular in the field of tumor biology in order to study cell-fate decisions or to trace cancer cells in the mouse. However, certain biological questions require a nongenetic labeling approach to purify a distinct cell population in the pancreas. Here we describe a protocol for isolating mouse pancreatic ductal epithelial cells and ductlike cells directly in vivo using ductal-specific Dolichos biflorus agglutinin (DBA) lectin labeling followed by magnetic bead separation. Isolated cells can be cultured (in two or three dimensions), manipulated by lentiviral transduction to modulate gene expression and directly used for molecular studies. This approach is fast (4 h), affordable, results in cells with high viability, can be performed on the bench and is applicable to virtually all genetic and nongenetic disease models of the pancreas.

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Figure 1: DBA lectin distribution in the normal pancreas and disease.
Figure 2
Figure 3: In vitro Cre recombination.
Figure 4: Ratio of relative gene expression in DBA lectin–positive and DBA lectin–negative cells.
Figure 5: Cells in 2D culture.
Figure 6: Confocal photomicrograph of cysts.
Figure 7: In vitro branching morphogenesis.
Figure 8: Cells in 3D culture.


  1. Gu, G., Dubauskaite, J. & Melton, D.A. Direct evidence for the pancreatic lineage: NGN3+ cells are islet progenitors and are distinct from duct progenitors. Development 129, 2447–2457 (2002).

    Article  CAS  Google Scholar 

  2. Kawaguchi, Y. et al. The role of the transcriptional regulator Ptf1a in converting intestinal to pancreatic progenitors. Nat. Genet. 32, 128–134 (2002).

    Article  CAS  Google Scholar 

  3. Reichert, M. & Rustgi, A.K. Pancreatic ductal cells in development, regeneration, and neoplasia. J. Clin. Invest. 121, 4572–4578 (2011).

    Article  CAS  Google Scholar 

  4. Desai, B.M. et al. Preexisting pancreatic acinar cells contribute to acinar cell, but not islet beta cell, regeneration. J. Clin. Invest. 117, 971–977 (2007).

    Article  CAS  Google Scholar 

  5. Solar, M. et al. Pancreatic exocrine duct cells give rise to insulin-producing beta cells during embryogenesis but not after birth. Dev. Cell 17, 849–860 (2009).

    Article  CAS  Google Scholar 

  6. Rhim, A.D. et al. EMT and dissemination precede pancreatic tumor formation. Cell 148, 349–361 (2012).

    Article  CAS  Google Scholar 

  7. Rovira, M. et al. Isolation and characterization of centroacinar/terminal ductal progenitor cells in adult mouse pancreas. Proc. Natl. Acad. Sci. USA 107, 75–80 (2010).

    Article  CAS  Google Scholar 

  8. Oshima, Y. et al. Isolation of mouse pancreatic ductal progenitor cells expressing CD133 and c-Met by flow cytometric cell sorting. Gastroenterology 132, 720–732 (2007).

    Article  CAS  Google Scholar 

  9. Agbunag, C., Lee, K.E., Buontempo, S. & Bar-Sagi, D. Pancreatic duct epithelial cell isolation and cultivation in two-dimensional and three-dimensional culture systems. Methods Enzymol. 407, 703–710 (2006).

    Article  CAS  Google Scholar 

  10. Gmyr, V. et al. Human pancreatic ductal cells: large-scale isolation and expansion. Cell Transplant. 10, 109–121 (2001).

    Article  CAS  Google Scholar 

  11. Cotton, C.U., Cotton, U. & Al-nakkash, L. Isolation and culture of bovine pancreatic duct epithelial cells Isolation and culture of bovine pancreatic duct epithelial cells. Am. J. Physiol. 272 (6 Part 1): G1328–G1337 (1997).

    CAS  PubMed  Google Scholar 

  12. Reichert, M. et al. The Prrx1 homeodomain transcription factor plays a central role in pancreatic regeneration and carcinogenesis. Genes Dev. 27, 288–300 (2013).

    Article  CAS  Google Scholar 

  13. Prasad, N.B. et al. Gene expression profiles in pancreatic intraepithelial neoplasia reflect the effects of Hedgehog signaling on pancreatic ductal epithelial cells. Cancer Res. 65, 1619–1626 (2005).

    Article  CAS  Google Scholar 

  14. Wang, Y.J., Bailey, J.M., Rovira, M. & Leach, S.D. Sphere-forming assays for assessment of benign and malignant pancreatic stem cells. Methods Mol. Biol. 980, 281–290 (2013).

    Article  CAS  Google Scholar 

  15. Hermann, P. et al. Distinct populations of cancer stem cells determine tumor growth and metastatic activity in human pancreatic cancer. Cell Stem Cell 1, 313–323 (2007).

    Article  CAS  Google Scholar 

  16. Li, C. et al. Identification of pancreatic cancer stem cells. Cancer Research 67, 1030–1037 (2007).

    Article  CAS  Google Scholar 

  17. Li, C. et al. c-Met is a marker of pancreatic cancer stem cells and therapeutic target. Gastroenterology 141, 2218–2227 e5 (2011).

    Article  CAS  Google Scholar 

  18. Cross, S.E. et al. AFM-based analysis of human metastatic cancer cells. Nanotechnology 19, 384003 (2008).

    Article  Google Scholar 

  19. Schreiber, F.S. et al. Successful growth and characterization of mouse pancreatic ductal cells: functional properties of the Ki-RASG12V oncogene. Gastroenterology 127, 250–260 (2004).

    Article  CAS  Google Scholar 

  20. Botta, G.P., Reginato, M.J., Reichert, M., Rustgi, A.K. & Lelkes, P.I. Constitutive K-RasG12D activation of ERK2 specifically regulates 3D invasion of human pancreatic cancer cells via MMP-1. Mol. Cancer Res. 10, 183–196 (2012).

    Article  CAS  Google Scholar 

  21. Wescott, M.P. et al. Pancreatic ductal morphogenesis and the Pdx1 homeodomain transcription factor. Mol. Biol. Cell 20, 4838–4844 (2009).

    Article  CAS  Google Scholar 

  22. Deramaudt, T.B. et al. N-cadherin and keratinocyte growth factor receptor mediate the functional interplay between Ki-RASG12V and p53V143A in promoting pancreatic cell migration, invasion, and tissue architecture disruption. Mol. Cell Biol. 26, 4185–4200 (2006).

    Article  CAS  Google Scholar 

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This work was supported by the US National Institutes of Health (NIH) (grant no. DK060694 to M.R., S.T., S.H., B.B., A.K.R.), the National Pancreas Foundation (M.R.), the Honjo International Scholarship Foundation (S.T.), Deutsche Krebshilfe (S.H.), the NIH/National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) (grant no. F30 DK088402-01 to G.P.B.), NIH/NIDDK grant no. P30 DK050306 Center for Molecular Studies in Digestive and Liver Diseases (and Molecular Pathology and Imaging, Molecular Biology/Gene Expression, Cell Culture, Transgenic and Chimeric Mouse Cores) and American Cancer Society grant no. RP-10-033-01-CCE.

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M.R. and A.K.R. conceived the DBA lectin sorting strategy. M.R., S.T., S.H., B.B., G.P.B. and A.K.R. performed the experiments and wrote the manuscript. M.R., S.T., S.H., B.B., G.P.B. and A.K.R. discussed the results and commented on the manuscript at all stages.

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Correspondence to Anil K Rustgi.

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The authors declare no competing financial interests.

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Reichert, M., Takano, S., Heeg, S. et al. Isolation, culture and genetic manipulation of mouse pancreatic ductal cells. Nat Protoc 8, 1354–1365 (2013).

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