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Replication of adult pancreatic-beta cells cultured on bovine corneal endothelial cell extracellular matrix

  • Growth, Differentiation And Senescence
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Summary

A valuable system has been developed to study replication of adult beta cells. Isolated islets from adult rats were partially dispersed and plated on dishes coated with extracellular matrix from bovine-corneal endothelial cells. Within 24 h islet cells attached to the matrix and formed a monolayer. The proportion of insulin-, glucagon-, and somatostatin-containing cells in the cultures was characteristic of whole islets. Function of beta cells was assessed by measuring glucosestimulated insulin release. Insulin release from 7-day-old cultures increased 19-fold after a 16.7 mM glucose challenge indicating that beta-cell function was normal. Cellular replication in the cultures was assessed using the thymidine analogue 5-bromo-2′-deoxyuridine (BrdU). BrdU incorporation was noted in insulin-, glucagon-, and somatostatin-containing cells and also in non-endocrine cells. Among endocrine cells, the majority of BrdU labeling occurred in beta cells. Beta-cell replication potential was assessed using different concentrations of glucose. The incorporation of BrdU into beta cells was affected in a dose-dependent manner by glucose; over a 10-fold increase of beta-cell BrdU labeling was observed when glucose concentration was raised from 5.5 to 16.7 mM. The system proved advantageous for studying the replication potential of adult beta cells.

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Authors and Affiliations

  1. Joslin Diabetes Center, New England Deaconess Hospital, Brigham

    G. T. Schuppin, S. Bonner-Weir, E. Montana, N. Kaiser & G. C. Weir

  2. Woman’s Hospital, and Harvard Medical School, 92215, Boston, Massachusetts

    G. T. Schuppin, S. Bonner-Weir, E. Montana, N. Kaiser & G. C. Weir

  3. Department of Endocrinology and Metabolism, Hebrew University Hadassah Medical Center, Jerusalem, Israel

    N. Kaiser

Authors
  1. G. T. Schuppin
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  2. S. Bonner-Weir
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  3. E. Montana
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  4. N. Kaiser
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  5. G. C. Weir
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Schuppin, G.T., Bonner-Weir, S., Montana, E. et al. Replication of adult pancreatic-beta cells cultured on bovine corneal endothelial cell extracellular matrix. In Vitro Cell Dev Biol - Animal 29, 339–344 (1993). https://doi.org/10.1007/BF02633963

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  • DOI: https://doi.org/10.1007/BF02633963

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