Isolated mouse pancreatic islets in culture: Effects of serum and different culture media on the insulin production of the islets

Summary

Various conditions for tissue culture of collagenase-isolated mouse pancreatic islets were studied in an attempt to optimize the maintenance of glucose stimulated insulin biosynthesis and release in the cultured specimens. Islets which had been cultured at a physiological glucose concentration (5.5 mmol/l) in the absence of serum had an impaired glucose-stimulated insulin biosynthesis and release as well as a reduced insulin content. Thus, insulin biosynthesis was three times higher after culture in a serum supplemented medium. Further, the insulin secretion of islets cultured in the presence of serum was markedly enhanced in acute incubations with high concentrations of glucose. This response was most pronounced in islets which had been cultured free-floating. A comparison between different culture media showed that islets cultured in RPMI 1640 had the highest insulin production. The present data suggest that the most favourable conditions for long-term storage of isolated islets in culture may be obtained when the islets are maintained as free-floating explants in a culture medium consisting of RPMI 1640 supplemented with serum.

References

  1. 1.

    Andersson, A.: Effects of glucose on the structure and metabolism of isolated pancreatic islets maintained in tissue culture. Thesis Acta Universitatis Upsalienses no. 169. Uppsala, Sweden, 1973

  2. 2.

    Andersson, A.: Synthesis of DNA in isolated pancreatic islets maintained in tissue culture. Endocrinology96, 1051–1054 (1975)

    PubMed  Google Scholar 

  3. 3.

    Andersson, A.: Tissue culture of isolated pancreatic islets. Acta Endocrinol. (Kbh.) [Suppl.]205, 283–293 (1976)

    Google Scholar 

  4. 4.

    Andersson, A., Hellerström, C.: Metabolic characteristics of isolated pancreatic islets in tissue culture. Diabetes21 [Suppl. 2], 546–554 (1972)

    PubMed  Google Scholar 

  5. 5.

    Andersson, A., Borg, H., Groth, C. G., Gunnarsson, R., Hellerström, C., Lundgren, G., Westman, J., Östman, J.: Survival of isolated human islets of Langerhans maintained in tissue culture. J. Clin. Invest.57, 1295–1301 (1976)

    PubMed  Google Scholar 

  6. 6.

    Andersson, A., Groth, C. G., Gunnarsson, R., Hellerström, C., Lundgren, G., Petersson, B., Östman, J.: Metabolic and secretory characteristics of cultured human islets of Langerhans. In: Proceedings of the 11th FEBS Meeting, Copenhagen, August 1977. New York: Pergamon Press42, 249–257 (1977)

    Google Scholar 

  7. 7.

    Andersson, A., Höiriis-Nielsen, J., Borg, H.: Effects of L-leucine on the insulin production, oxidative metabolism and mitochondrial ultrastructure of isolated mouse pancreatic islets in tissue culture. Diabetologia13, 59–69 (1977)

    PubMed  Google Scholar 

  8. 8.

    Berne, C.: Anti-insulin serum coupled to Sepharose 4B as a tool for the investigation of insulin biosynthesis in the B-cells of obese hyper-glycemic mice. Endocrinology97, 1241–1247 (1975)

    PubMed  Google Scholar 

  9. 9.

    Buitrago, A., Gylfe, E., Hellman, B., Idahl, L.-Å., Johansson, M.: Function of microdissected islets cultured in a chemically defined medium. I. Insulin content and release. Diabetologia11, 535–540 (1975)

    PubMed  Google Scholar 

  10. 10.

    Chick, W. L., Lauris, V., Flewelling, J. H., Andrews, K. A., Woodruff, J.: Effects of glucose on beta cells in pancreatic monolayer cultures. Endocrinology92, 212–218 (1973)

    PubMed  Google Scholar 

  11. 11.

    Eagle, H.: Amino acid metabolism in mammalian cell cultures. Science130, 432–437 (1959)

    PubMed  Google Scholar 

  12. 12.

    Elias, J. J., Riviera, E.: Comparison of the responses of normal, precancerous, and neoplastic mouse mammary tissues to hormones in vitro. Cancer Res.19, 505–511 (1959)

    PubMed  Google Scholar 

  13. 13.

    Frankel, B. J., Gylfe, E., Hellman, B., Idahl, L.-Å.: Maintenance of insulin release from pancreatic islets stored in the cold for up to 5 weeks. J. Clin. Invest.57, 47–52 (1976)

    PubMed  Google Scholar 

  14. 14.

    Goldman, H., Colle, E.: Human pancreatic islets in culture: Effects of supplementing the medium with homologous and heterologous serum. Science192, 1014–1016 (1976)

    PubMed  Google Scholar 

  15. 15.

    Heding, L.: Determination of total serum insulin (IRI) in insulin-treated diabetic patients. Diabetologia8, 260–266 (1972)

    PubMed  Google Scholar 

  16. 16.

    Holley, R. W., Kiernan, J. A.: Control of the initiation of DNA synthesis in 3T3 cells: Serum factors. Proc. Natl. Acad. Sci. U. S. A.71, 2908–2911 (1974)

    PubMed  Google Scholar 

  17. 17.

    Howell, S. L., Taylor, K. W.: Potassium ions and the secretion of insulin by islets of Langerhans incubated in vitro. Biochem. J.108, 17–24 (1968)

    PubMed  Google Scholar 

  18. 18.

    Kedinger, M., Moody, A., Lannay, J. F., Haffen, K.: Establishment of serum based medium essentially free of proteolytic activity for the culture of mouse pancreatic islets. Experientia33, 972–973 (1977)

    PubMed  Google Scholar 

  19. 19.

    Keen, H., Field, J. B., Pastan, I. H.: A simple method for in vitro metabolic studies using small volumes of tissue and medium. Metabolism12, 143–174 (1963)

    PubMed  Google Scholar 

  20. 20.

    Krebs, H. A., Henseleit, K.: Untersuchungen über die Harnstoffbildung im Tierkörper. Hoppe Seylers Z. Physiol. Chem.210, 33–66 (1932)

    Google Scholar 

  21. 21.

    Leach, F. N., Ashworth, M. A., Barson, A. J., Milner, R. D. G.: Insulin release from human foetal pancreas in tissue culture. J. Endocrinol.59, 65–79 (1973)

    PubMed  Google Scholar 

  22. 22.

    Matas, A. J., Sutherland, D. E. R., Najarian, J. S.: Current status of islet and pancreas transplantation in diabetes. Diabetes25, 785–795 (1976)

    PubMed  Google Scholar 

  23. 23.

    Morton, H. J.: A survey of commercially available tissue culture media. In Vitro6, 89–108 (1970)

    PubMed  Google Scholar 

  24. 24.

    Nielsen, J. H., Eff, C., Deckert, T., Peterssen, J., Brunstedt, K., Andersson, A.: Effect of diabetic sera on isolated islets in tissue culture. Diabetologia13, 421 (1977)

    Google Scholar 

  25. 25.

    Temin, H. M.: Studies on carcinogenesis by avian sarcoma viruses VI. Differential multiplication of uninfected and of converted cells in response to insulin. J. Cell Physiol.69, 377–384 (1967)

    PubMed  Google Scholar 

  26. 26.

    Täljedal, I.-B., Lernmark, L.-Å., Sehlin, J.: B-cytotoxiceffects of normal serum. Diabetologia13, 434 (1977)

    Google Scholar 

  27. 27.

    Ziegler, B., Hahn, H.-J., Ziegler, M., Fiedler, H.: Successful cultivation of isolated islets of Langerhans without attachment: Relationship between glucose- and theophyllineinduced insulin release and insulin content in rat islets after cultivation. Endokrinologie69, 103–111 (1977)

    PubMed  Google Scholar 

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Andersson, A. Isolated mouse pancreatic islets in culture: Effects of serum and different culture media on the insulin production of the islets. Diabetologia 14, 397–404 (1978). https://doi.org/10.1007/BF01228134

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Key words

  • Pancreatic islets
  • tissue culture
  • calf serum
  • culture media
  • insulin secretion
  • insulin content
  • insulin biosynthesis