Using Pancreas Tissue Slices for the Study of Islet Physiology

  • Julia K. Panzer
  • Christian M. Cohrs
  • Stephan SpeierEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2128)


Studies on islet of Langerhans physiology are crucial to understand the role of the endocrine pancreas in diabetes pathogenesis and the development of new therapeutic approaches. However, so far most research addressing islet of Langerhans biology relies on islets obtained via enzymatic isolation from the pancreas, which is known to cause mechanical and chemical stress, thus having a major impact on islet cell physiology. To circumvent the limitations of islet isolation, we have pioneered a platform for the study of islet physiology using the pancreas tissue slice technique. This approach allows to explore the detailed three-dimensional morphology of intact pancreatic tissue at a cellular level and to investigate islet cell function under near-physiological conditions. The described procedure is less damaging and faster than alternative approaches and particularly advantageous for studying infiltrated and structurally damaged islets. Furthermore, pancreas tissue slices have proven valuable for acute studies of endocrine as well as exocrine cell physiology in their conserved natural environment. We here provide a detailed protocol for the preparation of mouse pancreas tissue slices, the assessment of slice viability, and the study of pancreas cell physiology by hormone secretion and immunofluorescence staining.

Key words

Tissue slices In situ islet cell function Insulin secretion Immunofluorescence staining Live/dead staining 



This work was supported with funds from the Paul Langerhans Institute Dresden (PLID) of Helmholtz Zentrum München at the University Clinic Carl Gustav Carus of Technische Universität Dresden, the German Ministry of Education and Research (BMBF) to the German Centre for Diabetes Research (DZD), the DFG SFB/Transregio 127, the European Foundation for the Study of Diabetes (EFSD)/Boehringer Ingelheim Basic Research Programme, and the Helmsley Charitable Trust George S. Eisenbarth nPOD Award for Team Science.


  1. 1.
    Bhagat L et al (2000) Heat shock protein 70 prevents secretagogue-induced cell injury in the pancreas by preventing intracellular trypsinogen activation. J Clin Invest 106:81–89CrossRefGoogle Scholar
  2. 2.
    Blinman TA et al (2000) Activation of pancreatic acinar cells on isolation from tissue: cytokine upregulation via p38 MAP kinase. Am J Physiol Cell Physiol 279:C1993–C2003CrossRefGoogle Scholar
  3. 3.
    Irving-Rodgers HF et al (2012) Pancreatic islet basement membrane loss and remodeling after mouse islet isolation and transplantation: impact for allograft rejection. Cell Transplant 23:59–72CrossRefGoogle Scholar
  4. 4.
    Negi S et al (2012) Analysis of beta cell gene expression reveals inflammatory signaling and evidence of dedifferentiation following human islet isolation and culture. PLoS One 7:e30415CrossRefGoogle Scholar
  5. 5.
    Paraskevas S et al (1999) Activation and expression of ERK, JNK, and p38 MAP-kinases in isolated islets of Langerhans: implications for cultured islet survival. FEBS Lett 455:203–208CrossRefGoogle Scholar
  6. 6.
    Raposo do Amaral AS et al (2013) Glutathione ethyl ester supplementation during pancreatic islet isolation improves viability and transplant outcomes in a murine marginal islet mass model. PLoS One 8:e55288CrossRefGoogle Scholar
  7. 7.
    Speier S, Rupnik M (2003) A novel approach to in situ characterization of pancreatic beta-cells. Pflugers Arch 446:553–558CrossRefGoogle Scholar
  8. 8.
    Lacy PE, Kostianovsky M (1967) Method for the isolation of intact islets of Langerhans from the rat pancreas. Diabetes 16:35–39CrossRefGoogle Scholar
  9. 9.
    Li DS, Yuan YH, Tu HJ, Liang QL, Dai LJ (2009) A protocol for islet isolation from mouse pancreas. Nat Protoc 4:1649–1652CrossRefGoogle Scholar
  10. 10.
    Logsdon CD, Williams JA (1983) Pancreatic acini in short-term culture: regulation by EGF, carbachol, insulin, and corticosterone. Am J Phys 244:G675–G682Google Scholar
  11. 11.
    Ivanova A et al (2013) Age-dependent labeling and imaging of insulin secretory granules. Diabetes 62(11):3687–3696CrossRefGoogle Scholar
  12. 12.
    Marciniak A, Selck C, Friedrich B, Speier S (2013) Mouse pancreas tissue slice culture facilitates long-term studies of exocrine and endocrine cell physiology in situ. PLoS One 8:e78706CrossRefGoogle Scholar
  13. 13.
    Marciniak A, Cohrs C et al (2014) Using pancreas tissue slices for in situ studies of islet of Langerhans and acinar cell biology. Nat Protoc 9(12):2809–2822CrossRefGoogle Scholar
  14. 14.
    Speier S, Yang SB, Sroka K, Rose T, Rupnik M (2005) KATP-channels in beta cells in tissue slices are directly modulated by millimolar ATP. Mol Cell Endocrinol 230:51–58CrossRefGoogle Scholar
  15. 15.
    Speier S, Gjinovci A, Charollais A, Meda P, Rupnik M (2007) Cx36-mediated coupling reduces beta cell heterogeneity, confines the stimulating glucose concentration range, and affects insulin release kinetics. Diabetes 56:1078–1086CrossRefGoogle Scholar
  16. 16.
    Huang YC et al (2013) In situ electrophysiological examination of pancreatic alpha cells in the streptozotocin-induced diabetes model, revealing the cellular basis of glucagon hypersecretion. Diabetes 62:519–530CrossRefGoogle Scholar
  17. 17.
    Rose T, Efendic S, Rupnik M (2007) Ca2+−secretion coupling is impaired in diabetic Goto Kakizaki rats. J Gen Physiol 129:493–508CrossRefGoogle Scholar
  18. 18.
    Huang YC, Rupnik M, Gaisano HY (2011) Unperturbed islet alpha cell function examined in mouse pancreas tissue slices. J Physiol 589:395–408CrossRefGoogle Scholar
  19. 19.
    Cohrs C, Chen C et al (2017) Vessel network architecture of adult human islets promotes distinct cell-cell interactions in situ and is altered after transplantation. Endocrinology 158(5):1373–1385CrossRefGoogle Scholar
  20. 20.
    Weitz J et al (2018) Mouse pancreatic islet macrophages use locally released ATP to monitor beta cell activity. Diabetologia 61(1): 182–192CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Julia K. Panzer
    • 1
    • 2
    • 3
  • Christian M. Cohrs
    • 1
    • 2
    • 3
  • Stephan Speier
    • 1
    • 2
    • 3
    Email author
  1. 1.Paul Langerhans Institute Dresden (PLID) of the Helmholtz Zentrum München at the University Clinic Carl Gustav Carus of Technische Universität Dresden, Helmholtz Zentrum MünchenNeuherbergGermany
  2. 2.Institute of Physiology, Faculty of Medicine, Technische Universität DresdenDresdenGermany
  3. 3.German Center for Diabetes Research (DZD)München-NeuherbergGermany

Personalised recommendations