Abstract
Islets of Langerhans isolated from experimental animals, such as mice and rats, have contributed much to our understanding of the mechanisms by which pancreatic β-cells secrete insulin in a regulated manner, and this knowledge is important in identifying potential novel therapies for Type 2 diabetes. However, although many of the signal transduction pathways identified in rodent islets are common to humans, some critical differences have been demonstrated experimentally. It is, therefore, essential that experiments are performed using islets isolated from human pancreas to provide robust data defining whether the key observations made in rodents are also applicable to the human situation. The rate-limiting factor in this area of research is the supply of high-quality human islets isolated from pancreases retrieved from organ donors, and the protocols described in this chapter aim to provide methods of maximising information that can be obtained when human islets are maintained in culture.
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References
Cabrera, O., Berman, D.M., Kenyon, N.S., Ricordi, C., Berggren, P.O. and Caicedo, A. (2006) The unique cytoarchitecture of human pancreatic islets has implications for islet cell function. Proc Natl Acad Sci USA. 103, 2334–9.
Kim, A., Miller, K., Jo, J., Kilimnik, G., Wojcik, P. and Hara, M. (2009) Islet architecture. A comparative study. Islets 1, 129–136.
Davalli, A.M.,Biancardi, E., Pollo, A., Socci, C., Pontiroli, A.E., Pozza, G., Clementi, F., Sher, E. and Carbone, E. (1996) Dihydropyridine-sensitive and -insensitive voltage-operated calcium channels participate in the control of glucose-induced secretion from human pancreatic beta cells. J.Endocrinol. 150, 195–203.
Martin, F. and Soria, B. (1996) Glucose-induced (Ca2+) oscillations in single human pancreatic islets. Cell Calcium 20, 409–414.
Huang, G.C., Zhao, M., Jones, P.M., Persaud, S., Ramracheya, R., Lobner, K., et al. (2004) The development of new density gradient media for the purification of human islets and islet quality assessments. Transplantation 77, 143–145.
Varadi, A., Molnar, E. and Ashcroft, S.J.H. (1996) A unique combination of plasma membrane Ca2+/ATPase isoforms is expressed in islets of Langerhans and pancreatic β-cell lines. Biochem. J. 314, 663–669.
Lacey, R.J., Berrow, N.S., London, N.J.M., Lake, S.P., James, R.F.L., Scarpello, J.H.B. and Morgan, N.G. (1990) Differential effects of β-adrenergic agonists on insulin secretion from pancreatic islets isolated from rat and man. J. Mol. Endocrinol. 5, 49–54.
Li, G., Milani, D., Dunne, M.J., Pralong, W., Theler, J., Petersen, O.H. and Wollheim, C.B. (1991) Extracellular ATP causes Ca2+-dependent and -independent insulin secretion in RINm5F cells. J. Biol. Chem. 266, 3449–3452.
Squires, P.E., James, R.F.L., London, N.J.M. and Dunne, M.J. (1994) ATP-induced intracellular Ca2+ signals in isolated human insulin-secreting cells. Pflugers Arch. 427, 181–183.
Ramracheya, R.D., Muller, D., Squires, P.E., Brereton, H., Sugden, D., Huang, G.C. Amiel, S.A., Jones, P.M. and Persaud, S.J. (2008) Function and expression of melatonin receptors on human pancreatic islets. J. Pineal Res. 44,273–279.
Muller, D., Jones, P.M. and Persaud, S.J. (2008) Expression of a potent anti-apoptotic protein in MIN6 cells using METAFECTENE PRO, Application note. http://www.biontex.com/con_4_6_4/cms/upload/pdf/Muller_MP_en.pdf.
Muller, D., Huang, G.C., Amiel, S., Jones, P.M. and Persaud, S.J. (2006) Identification of insulin signalling elements in human β-cells: autocrine regulation of insulin gene expression. Diabetes 55, 2835–2842.
Acknowledgements
The authors are grateful to the relatives of organ donors for human pancreases and to Professor S. Amiel and Dr. G.C. Huang for provision of isolated human islets.
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Persaud, S.J., Liu, B., Jones, P.M. (2012). Functional Analysis of Human Islets of Langerhans Maintained in Culture. In: Mitry, R., Hughes, R. (eds) Human Cell Culture Protocols. Methods in Molecular Biology, vol 806. Humana Press. https://doi.org/10.1007/978-1-61779-367-7_5
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DOI: https://doi.org/10.1007/978-1-61779-367-7_5
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