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Using pancreas tissue slices for in situ studies of islet of Langerhans and acinar cell biology

Nature Protocols volume 9, pages 2809–2822 (2014)Cite this article

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Abstract

Studies on the cellular function of the pancreas are typically performed in vitro on its isolated functional units, the endocrine islets of Langerhans and the exocrine acini. However, these approaches are hampered by preparation-induced changes of cell physiology and the lack of an intact surrounding. We present here a detailed protocol for the preparation of pancreas tissue slices. This procedure is less damaging to the tissue and faster than alternative approaches, and it enables the in situ study of pancreatic endocrine and exocrine cell physiology in a conserved environment. Pancreas tissue slices facilitate the investigation of cellular mechanisms underlying the function, pathology and interaction of the endocrine and exocrine components of the pancreas. We provide examples for several experimental applications of pancreas tissue slices to study various aspects of pancreas cell biology. Furthermore, we describe the preparation of human and porcine pancreas tissue slices for the validation and translation of research findings obtained in the mouse model. Preparation of pancreas tissue slices according to the protocol described here takes less than 45 min from tissue preparation to receipt of the first slices.

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Figure 1: Mouse tissue preparation and vibratome slicing.
Figure 2: Live-cell morphology of in situ pancreas tissue.
Figure 3: 3D morphology of mouse pancreas tissue after antibody staining.
Figure 4: Antibody staining of exocrine tissue in mouse pancreas slices.
Figure 5: Calcium imaging of beta cells within a mouse pancreas tissue slice, using a genetically encoded calcium indicator.
Figure 6: Human pancreas tissue slices.
Figure 7: Porcine pancreas tissue slices.
Figure 8
Figure 9

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Acknowledgements

This work was supported with funds from the Emmy Noether Program of the German Research Foundation (DFG), the CRTD—DFG Research Center for Regenerative Therapies Dresden, Cluster of Excellence, the DFG-SFB/Transregio 127 and the German Ministry for Education and Research (BMBF) to the German Centre for Diabetes Research (DZD) and to the Network of Competence for Diabetes (KKNDm). The work leading to this publication has received support from the Innovative Medicines Initiative Joint Undertaking under grant agreement no. 155005 ('improving beta cell function and identification of diagnostic biomarkers for treatment monitoring in diabetes'; IMIDIA), resources of which are composed of financial contribution from the European Union's Seventh Framework Programme (FP7/2007-2013) and the European Federation of Pharmaceutical Industries and Associations (EFPIA) companies' in-kind contributions. We thank K. Hüttner, C. Kühn, A. Gröbe and K. Pfützner for their excellent technical assistance, and D. Richter, M. Roessler, M. Distler, R. Grutzmann and G. Baretton for providing human pancreatic tissues.

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Author notes

  1. Anja Marciniak and Christian M Cohrs: These authors contributed equally to this work.

Authors and Affiliations

  1. Deutsche Forschungsgemeinschaft (DFG) Research Center for Regenerative Therapies Dresden (CRTD), Technische Universität Dresden, Dresden, Germany.,

    Anja Marciniak, Christian M Cohrs, Vasiliki Tsata, Julie A Chouinard, Claudia Selck, Julia Stertmann, Saskia Reichelt & Stephan Speier

  2. Paul Langerhans Institute Dresden, University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany

    Anja Marciniak, Christian M Cohrs, Vasiliki Tsata, Julie A Chouinard, Claudia Selck, Julia Stertmann, Saskia Reichelt, Michele Solimena & Stephan Speier

  3. German Center for Diabetes Research (DZD), Dresden, Germany.,

    Anja Marciniak, Christian M Cohrs, Vasiliki Tsata, Julie A Chouinard, Claudia Selck, Julia Stertmann, Saskia Reichelt, Michele Solimena & Stephan Speier

  4. Max Planck Institute of Neurobiology, Martinsried, Germany

    Tobias Rose

  5. Department of GI-, Thoracic- and Vascular Surgery, University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany.,

    Florian Ehehalt & Jürgen Weitz

  6. Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany

    Michele Solimena

  7. Institute of Physiology, Faculty of Medicine, University of Maribor, Maribor, Slovenia

    Marjan Slak Rupnik

  8. Institute of Physiology, Center for Physiology and Pharmacology, Medical University of Vienna, Vienna, Austria

    Marjan Slak Rupnik

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  1. Anja Marciniak
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Contributions

M.S.R. and S.S. developed the original tissue slice preparation for mouse pancreas; A.M., C.M.C., V.T., J.A.C., C.S., J.S., S.R. and S.S. developed the organotypic culture of adult mouse pancreas slices, the perifusion of slices for calcium imaging, the secretion assays and immunohistochemistry protocols for slices, and the slicing procedure for human and porcine pancreas; T.R. and M.S.R. adapted the tissue slice preparation for rat pancreas; F.E., J.W. and M.S. provided living human pancreas tissue; and A.M., C.M.C., M.S.R. and S.S. wrote the manuscript.

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Correspondence to Stephan Speier.

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Marciniak, A., Cohrs, C., Tsata, V. et al. Using pancreas tissue slices for in situ studies of islet of Langerhans and acinar cell biology. Nat Protoc 9, 2809–2822 (2014). https://doi.org/10.1038/nprot.2014.195

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