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. 1976 May;57(5):1295–1301. doi: 10.1172/JCI108397

Survival of isolated human islets of Langerhans maintained in tissue culture.

A Andersson, H Borg, C G Groth, R Gunnarsson, C Hellerström, G Lundgren, J Westman, J Ostman
PMCID: PMC436782  PMID: 770504

Abstract

Transplantation of human pancreatic islets to diabetic patients may require that donor islets be kept viable in vitro for extended time periods before transfer to the recipient. We have maintained isolated pancreatic islets obtained from the human cadaveric pancreas in tissue culture for 1-3 wk, after which we studied the structure and function of the islets. Electron micrographs of the cultured islets showed a satisfactory preservation of both beta-cells and alpha 2-cells. After culture for 1 wk, the islet oxygen uptake proceeded at a constant rate at a low glucose concentration (3.3 mM) and was significantly enhanced by raising the glucose concentration to 16.7 mM. Likewise, after culture for 1 wk, the islets responded with an increased insulin release when exposed to 16.7 mM glucose with or without added theophylline (10 mM). Islets cultured for 1-3 wk were able to incorporate [3H]leucine into proinsulin, as judged by gel filtration of acid-alcohol extracts. Glucagon release from the cultured islets was reduced significantly by 16.7 mM glucose alone, but stimulated by glucose (16.7 mM) plus theophylline (10 MM). It is concluded that viable pancreatic islets can be isolated from the pancreas of adult human donors and maintained in tissue culture for at least 1 wk without loss of the specific functions of the alpha 2- and beta-cells. It remains to be established whether such islets will survive and remain functionally competent after transplantation to human recipients.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Amamoo D. G., Woods J. E., Donovan J. L. Preliminary experience with pancreatic islet-cell implantation. Mayo Clin Proc. 1974 May;49(5):289–292. [PubMed] [Google Scholar]
  2. Andersson A., Hellerström C. Metabolic characteristics of isolated pancreatic islets in tissue culture. Diabetes. 1972;21(2 Suppl):546–554. doi: 10.2337/diab.21.2.s546. [DOI] [PubMed] [Google Scholar]
  3. Andersson A., Westman J., Hellerström C. Effects of glucose on the ultrastructure and insulin biosynthesis of isolated mouse pancreatic islets maintained in tissue culture. Diabetologia. 1974 Dec;10(6):743–753. doi: 10.1007/BF01219536. [DOI] [PubMed] [Google Scholar]
  4. Ashcroft S. J., Bassett J. M., Randle P. J. Isolation of human pancreatic islets capable of releasing insulin and metabolising glucose in vitro. Lancet. 1971 May 1;1(7705):888–889. doi: 10.1016/s0140-6736(71)92445-7. [DOI] [PubMed] [Google Scholar]
  5. Asplund K., Hellerström C. Glucose metabolism of pancreatic islets isolated from neonatal rats. Horm Metab Res. 1972 May;4(3):159–163. doi: 10.1055/s-0028-1094091. [DOI] [PubMed] [Google Scholar]
  6. Ballinger W. F., Lacy P. E. Transplantation of intact pancreatic islets in rats. Surgery. 1972 Aug;72(2):175–186. [PubMed] [Google Scholar]
  7. Barker C. F. Transplantation of the islets of Langerhans and the histocompatibility of endocrine tissue. Diabetes. 1975 Aug;24(8):766–775. doi: 10.2337/diab.24.8.766. [DOI] [PubMed] [Google Scholar]
  8. Chesney T. M., Schofield J. G. Studies on the secretion of pancreatic glucagon. Diabetes. 1969 Sep;18(9):627–632. doi: 10.2337/diab.18.9.627. [DOI] [PubMed] [Google Scholar]
  9. Clark J. L., Steiner D. F. Insulin biosynthesis in the rat: demonstration of two proinsulins. Proc Natl Acad Sci U S A. 1969 Jan;62(1):278–285. doi: 10.1073/pnas.62.1.278. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Connolly J. E., Martin D. C., Steinberg T., Gwinup G., Gazzaniga A. B., Bartlett R. H. Clinical experience with pancreaticoduodenal transplantation. Arch Surg. 1973 Apr;106(4):489–494. doi: 10.1001/archsurg.1973.01350160107017. [DOI] [PubMed] [Google Scholar]
  11. Creutzfeldt C., Track N. S., Creutzfeldt W. In vitro studies of the rate of proinsulin and insulin turnover in seven human insulinomas. Eur J Clin Invest. 1973 Sep;3(5):371–384. doi: 10.1111/j.1365-2362.1973.tb02203.x. [DOI] [PubMed] [Google Scholar]
  12. DAVOREN P. R. The isolation of insulin from a single cat pancreas. Biochim Biophys Acta. 1962 Sep 10;63:150–153. doi: 10.1016/0006-3002(62)90347-5. [DOI] [PubMed] [Google Scholar]
  13. Edwards J. C., Hellerström C., Petersson B., Taylor K. W. Oxidation of glucose and fatty acids in normal and in A 2 -cell rich pancreatic islets isolated from guinea-pigs. Diabetologia. 1972 Apr;8(2):93–98. doi: 10.1007/BF01235632. [DOI] [PubMed] [Google Scholar]
  14. Gliedman M. L., Gold M., Whittaker J., Rifkin H., Soberman R., Freed S., Tellis V., Veith F. J. Clinical segmental pancreatic transplantation with ureter-pancreatic duct anastomosis for exocrine drainage. Surgery. 1973 Aug;74(2):171–180. [PubMed] [Google Scholar]
  15. Goetz F. C. Conference on beta cell function, transplantation, and implantable glucose sensors: a summary. Metabolism. 1974 Sep;23(9):875–884. doi: 10.1016/0026-0495(74)90121-8. [DOI] [PubMed] [Google Scholar]
  16. HOLTER H. The Cartesian diver. Gen Cytochem Methods. 1961;2:93–129. doi: 10.1016/b978-0-12-395584-5.50007-x. [DOI] [PubMed] [Google Scholar]
  17. Hahn H. J., Ziegler M., Mohr E. Inhibition of glucagon secretion by glucose and glyceraldehyde on isolated islets of Wistar rats. FEBS Lett. 1974 Dec 1;49(1):100–102. doi: 10.1016/0014-5793(74)80641-1. [DOI] [PubMed] [Google Scholar]
  18. Hedeskov C. J., Hertz L., Nissen C. The effect of mannoheptulose on glucose- and pyruvate-stimulated oxygen uptake in normal mouse pancreatic islets. Biochim Biophys Acta. 1971 Feb 28;261(2):388–397. doi: 10.1016/0304-4165(72)90063-3. [DOI] [PubMed] [Google Scholar]
  19. Heding L. G. Radioimmunological determination of pancreatic and gut glucagon in plasma. Diabetologia. 1971 Feb;7(1):10–19. doi: 10.1007/BF02346248. [DOI] [PubMed] [Google Scholar]
  20. Howell S. L., Edwards J. C., Montague W. Regulation of adenylate cyclase and cyclic-AMP dependent protein kinase activities in A2-cell rich guinea-pig islets of Langerhans. Horm Metab Res. 1974 Jan;6(1):49–52. doi: 10.1055/s-0028-1093883. [DOI] [PubMed] [Google Scholar]
  21. Jarrousse C., Rosselin G. Interaction of amino acids and cyclic AMP on the release of insulin and glucagon by newborn rat pancreas. Endocrinology. 1975 Jan;96(1):168–177. doi: 10.1210/endo-96-1-168. [DOI] [PubMed] [Google Scholar]
  22. Kemp C. B., Knight M. J., Scharp D. W., Lacy P. E., Ballinger W. F. Transplantation of isolated pancreatic islets into the portal vein of diabetic rats. Nature. 1973 Aug 17;244(5416):447–447. doi: 10.1038/244447a0. [DOI] [PubMed] [Google Scholar]
  23. Lawson R. K., Poutala S. D. Tissue culture of adult human islets of Langerhans for transplantation. Surg Forum. 1974;25(0):377–379. [PubMed] [Google Scholar]
  24. Lillehei R. C., Simmons R. L., Najarian J. S., Weil R., Uchida H., Ruiz J. O., Kjellstrand C. M., Goetz F. C. Pancreatico-duodenal allotransplantation: experimental and clinical experience. Ann Surg. 1970 Sep;172(3):405–436. doi: 10.1097/00000658-197009000-00010. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Marliss E. B., Wollheim C. B., Blondel B., Orci L., Lambert A. E., Stauffacher W., Like A. A., Renold A. E. Insulin and glucagon release from monolayer cell cultures of pancreas from newborn rats. Eur J Clin Invest. 1973 Jan;3(1):16–26. doi: 10.1111/j.1365-2362.1973.tb00324.x. [DOI] [PubMed] [Google Scholar]
  26. Petersson B., Hellerström C., Gunnarsson R. Structure and metabolism of the pancreatic islets in streptozotocin treated guinea pigs. Horm Metab Res. 1970 Nov;2(6):313–317. doi: 10.1055/s-0028-1095064. [DOI] [PubMed] [Google Scholar]
  27. REYNOLDS E. S. The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J Cell Biol. 1963 Apr;17:208–212. doi: 10.1083/jcb.17.1.208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Reckard C. R., Barker C. F. Transplantation of isolated pancreatic islets across strong and weak histocompatibility barriers. Transplant Proc. 1973 Mar;5(1):761–763. [PubMed] [Google Scholar]
  29. Reckard C. R., Ziegler M. M., Naji A., Galbut D., Barker C. F. Physiologic and immunologic status of long-functioning transplanted pancreatic islets in rats. Surg Forum. 1974;25(0):374–376. [PubMed] [Google Scholar]
  30. Sasaki H., Rubalcava B., Baetens D., Blazquez E., Srikant C. B., Orci L., Unger R. H. Identification of glucagon in the gastrointestinal tract. J Clin Invest. 1975 Jul;56(1):135–145. doi: 10.1172/JCI108062. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Scharp D. W., Murphy J. J., Newton W. T., Ballinger W. F., Lacy P. E. Transplantation of islets of Langerhans in diabetic rhesus monkeys. Surgery. 1975 Jan;77(1):100–105. [PubMed] [Google Scholar]
  32. Steiner D. F., Oyer P. E. The biosynthesis of insulin and a probable precursor of insulin by a human islet cell adenoma. Proc Natl Acad Sci U S A. 1967 Feb;57(2):473–480. doi: 10.1073/pnas.57.2.473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Sutherland D. E., Steffes M. W., Bauer G. E., McManus D., Noe B. D., Najarian J. S. Isolation of human and porcine islets of Langerhans and islet transplantation in pigs. J Surg Res. 1974 Feb;16(2):102–111. doi: 10.1016/0022-4804(74)90017-1. [DOI] [PubMed] [Google Scholar]
  34. Vance J. E., Buchanan K. D., Challoner D. R., Williams R. H. Effect of glucose concentration on insulin and glucagon release from isolated islets of Langerhans of the rat. Diabetes. 1968 Apr;17(4):187–193. doi: 10.2337/diab.17.4.187. [DOI] [PubMed] [Google Scholar]
  35. WATSON M. L. Staining of tissue sections for electron microscopy with heavy metals. J Biophys Biochem Cytol. 1958 Jul 25;4(4):475–478. doi: 10.1083/jcb.4.4.475. [DOI] [PMC free article] [PubMed] [Google Scholar]

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