Diabetes, Vol 44, Issue 4 375-381, Copyright © 1995 by American Diabetes Association

ARTICLES

Massive isolation, morphological and functional characterization, and xenotransplantation of bovine pancreatic islets

P Marchetti, R Giannarelli, S Cosimi, P Masiello, A Coppelli, P Viacava and R Navalesi
Cattedra di Malattie del Metabolismo, Universita di Pisa, Italy.

The limited availability of human donors makes the search for alternative islet sources mandatory for future developments in pancreatic islet transplantation. In this study, we report on the massive isolation of bovine islets of proven in vitro and in vivo viability. The islets were prepared by collagenase digestion, sequential filtrations, and density-gradient purification by modifying a technique previously developed in our laboratory for the porcine pancreas. The prepurification yield was 2,743 +/- 78 islet equivalents (IE)/g pancreas (mean +/- SE), with a postpurification recovery of 78.7 +/- 2.2%. Purity ranged from 80 to 90%. The histological and immunocytochemical studies demonstrated the identity and integrity of the islets with well-preserved insulin-, glucagon-, and somatostatin-containing cells. The morphological integrity of cultured bovine islets was demonstrated for up to 4 weeks from isolation. Insulin secretion from freshly isolated islets was similar at 3.3 mmol/l glucose (0.36 +/- 0.06 pmol.IE-1.min-1) and at 14 mmol/l glucose (0.42 +/- 0.00 pmol.IE-1.min-1), and it increased significantly (P < 0.01) at 25 mmol/l glucose (1.44 +/- 0.12 pmol.IE-1.min-1). Arginine, theophylline, and propionic acid increased insulin secretion from freshly isolated islets at 3.3 and 14 mmol/l glucose, but not at 25 mmol/l glucose. Islets cultured at 37 degrees C in CMRL 1066 culture medium for at least 10 days were shown to become responsive to a lower glucose concentration, as demonstrated by the significant increase of insulin release in response to 14 mmol/l glucose, when compared with basal secretion. This recovered responsivity to glucose was maintained after 4 weeks of culture.(ABSTRACT TRUNCATED AT 250 WORDS)
CiteULike    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				P. Marchetti, A. Antonelli, R. Lupi, L. Marselli, P. Fallahi, C. Nesti, G. Baj, and E. Ferrannini Prolonged In Vitro Exposure to Autoantibodies Against CD38 Impairs the Function and Survival of Human Pancreatic Islets Diabetes, December 1, 2002; 51(90003): S474 - 477. [Abstract] [Full Text] [PDF]

				A. Antonelli, G. Baj, P. Marchetti, P. Fallahi, N. Surico, C. Pupilli, F. Malavasi, and E. Ferrannini Human Anti-CD38 Autoantibodies Raise Intracellular Calcium and Stimulate Insulin Release in Human Pancreatic Islets Diabetes, May 1, 2001; 50(5): 985 - 991. [Abstract] [Full Text]

				B. ROSATI, P. MARCHETTI, O. CROCIANI, M. LECCHI, R. LUPI, A. ARCANGELI, M. OLIVOTTO, and E. WANKE Glucose- and arginine-induced insulin secretion by human pancreatic {beta}-cells: the role of HERG K+ channels in firing and release FASEB J, December 1, 2000; 14(15): 2601 - 2610. [Abstract] [Full Text]