Summary
The roles of glucose and insulin in the promotion of DNA synthesis in pancreatic islet cell monolayers were assessed using a variety of in vitro conditions. Several substrates including collagen, poly-l-lysine, Matrigel, and the extracellular matrix produced by cultured bovine endothelial cells (BCEM) were compared for their ability to promote monolayer growth. Islets grown on BCEM in combination with medium RPMI 1640 supplemented with 22.2 mM glucose or 10 μg/ml insulin gave the best results as determined by new DNA synthesis. The new-form monolayers were free of contaminating, fibroblasts. These results suggest that insulin is critical to pancreatic islet growth when the cells are attached to biocompatible matrices.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
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.
Bottenstein, J.; Hayashi, I.; Utchings, S., et al. The growth of cells in serum-free hormone-supplemented media. In: Jakoby, W. B.; Pastan, I. M., eds. Methods in enzymology. New York: Academic Press; 1979:95–109.
Bowersox, J. C.; Sorgente, N. Altered growth kinetics of dermal fibroblasts and arterial smooth muscle cells from spontaneously diabetic BB rats. Diabetes 34:628–633; 1985.
Crickard, K.; Charles, R. I.; Jaffe, R. B. Control of proliferation of human fetal adrenal cells in vitro. J. Clin. Endocrinol. Metab. 53:790–796; 1981.
Eriksson, U.; Swenne, I. Diabetes in pregnancy: growth of the fetal pancreatic B-cells in the rat. Biol. Neonate 42:239–248; 1982.
Gospodarowicz, D.; Lepine, J.; Massoglia, S., et al. Comparison of the ability of basement membranes produced by corneal endothelial and mouse-derived endodermal PF-HR-9 cells to support the proliferation and differentations of bovine kidney tubule epithelial cells in vitro. J. Cell. Biol. 99:947–961; 1984.
Hayek, A.; Barela, T. D.; Wogenrich, F. J., et al. Fetal rat islet insulin deficiency following maternal administration of streptozotocin. Proc. Soc. Exp. Biol. Med. 180:209–213; 1985.
Hayek, A.; Guardian, C. Hormone release, islet yield, and transplantation of fetal and neonatal rat dorsal and ventral pancreatic islets. Diabetes 35:1189–1192; 1986.
Hayek, A.; Lopez, A. D.; Beattie, G. M. Decrease in number of neonatal islets required for successful transplantation by strict metabolic control of diabetic rats. Transplantation 45:940–942; 1988.
Hayek, A.; Woodside, W. Correlation between morphology and function in isolated islets of the Zucker rat. Diabetes 28:565–569; 1979.
Hellerstrom, C.; Lewis, N.; Borg, H., et al. Method for largescale isolation of pancreatic islets by tissue culture of fetal rat pancreas. Diabetes 28:767–776; 1979.
Hellerstrom, C.; Swenne, I. Growth pattern of pancreatic islets in animals. In: Volk, B. W.; Arquilla, E. R., eds. The diabetic pancreas. New York and London: Plenum Publishing; 1985:53–79.
Hinegardner, R. T. An improved fluorometric assay for DNA. Anal. Biochem. 39:197–201; 1971.
Kaiser, N.; Corcos, A. P.; Tur-Sinai, A., et al. Monolayer culture of adult rat pancreatic islets on extracellular matrix: long term maintenance of differentiated B-cell function. Endocrinology 123:834–840; 1988.
King, D. L.; Kyle, K. C.; Chick, W. L. Pancreatic B-cell replication: relation to insulin secretion. Endocrinology 103:1321–1327; 1978.
Loo, D. T.; Fuquay, J. I.; Rawson, C. L., et al. Extended culture of mouse embryo cells without senescence: inhibition by serum. Science 236:200–202; 1987.
Masquelier, D.; Amory, B.; Mourmeaux, J. L., et al. Cell interactions during in vitro neoformation of fetal rat pancreatic islets. Cell Differ. 18:199–211; 1986.
Mourmeauz, J. L.; Remacle, C.; Henquin, J. C. Morphological and functional characteristics of islets neoformed during tissue culture of fetal rat pancreas. Mol. Cell. Endocrinol. 39:237–246; 1985.
Ohgawara, H.; Carroll, R.; Hofmann, C., et al. Promotion of monolayer formation in cultured whole pancreatic islets by 3-isobutyl-l-methylxantahine. Proc. Natl. Acad. Sci. USA 75:1897–1900; 1978.
Rabinovitch, A.; Blondel, B.; Murray, T. Cyclic adenosine −3′,5′-monophosphate stimulates islet B-cell replication in neonatal rat pancreatic monolayer cultures. J. Clin. Invest. 66:1065–1071; 1980.
Rabinovitch, A.; Quigley, C.; Russell, T., et al. Insulin and multiplication stimulating activity (an insulin-like growth factor) stimulate islet B-cell replication in neonatal rat pancreatic monolayer cultures. Diabetes 31:160–164; 1982.
Shimizu, S.; Kagawa, S.; Nakao, K., et al. Effect of iodoacetic acid on maintenance of pancreatic endocrine cells in the neonatal rat. Endocrinol. Jpn. 31:253–261; 1984.
Swenne, I. The role of glucose in thein vitro regulation of cell cycle kinetics and proliferation of fetal pancreatic B-cells. Diabetes 31:754–760; 1982.
Swenne, I.; Bone, A. J.; Howell, S. L., et al. Effects of glucose and amino acids on the biosynthesis of DNA and insulin in fetal rat islets maintenance in tissue culture. Diabetes 29:686–692; 1980.
Swenne, I.; Eriksson, U. Diabetes in pregnancy: islet cell proliferation in the fetal rat pancreas. Diabetologia 23:525–528; 1982.
Thivolet, C. H.; Chatelain, P.; Nicoloso, H., et al. Morphological and functional effects of extracellular matrix on pancreatic islet cell culture. Exp. Cell Res. 159:313–322; 1985.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Hayek, A., Lopez, A.D. & Beattie, G.M. Enhancement of pancreatic islet cell monolayer growth by endothelial cell matrix and insulin. In Vitro Cell Dev Biol 25, 146–150 (1989). https://doi.org/10.1007/BF02626171
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02626171