Vol 59, No 4 (2000)
Original article
Submitted: 2012-02-06
Published online: 2000-08-02
Ultrastructural immunogold study on the various cell types of cultured pancreatic islets of adult rats
Mostafa M El-Naggar
Folia Morphol 2000;59(4):253-262.
Vol 59, No 4 (2000)
ORIGINAL ARTICLES
Submitted: 2012-02-06
Published online: 2000-08-02
Abstract
Whereas several reports describing the ultrastructure of the intact pancreatic islets have been recorded, published experience with the ultrastructural integrity of the cultured pancreatic islets is limited. The present study was, therefore, undertaken to provide an ultrastructure identification of the different cells in the cultured islets of the adult rat pancreas, after marking their secretory granules with gold particles. Pancreatic islets were isolated from adult male Wistar rats by the intraductal perfusion of collagenase technique. The islets were cultured in RPMI-1640 medium for 3 days and processed for preparation of ultrathin sections. The sections were stained with the indirect immunogold technique for insulin, glucagon, somatostatin, and pancreatic polypeptide. Ultrastructural examination of the cultured islets clearly identified the presence of B, A, D and PP-cells, as indicated by the numerous gold particles concentrated predominantly over the secretory granules. The secretory granules of the various cell types of the cultured islets demonstrated several similarities as well as differences from the recorded results of the corresponding secretory granules of the intact islets. The differences probably reflect a deviation in the underlying mechanisms of synthesis, maturation and secretion of the different secretory products of the cells in the cultured islets as they adapt to the in vitro environment.
Abstract
Whereas several reports describing the ultrastructure of the intact pancreatic islets have been recorded, published experience with the ultrastructural integrity of the cultured pancreatic islets is limited. The present study was, therefore, undertaken to provide an ultrastructure identification of the different cells in the cultured islets of the adult rat pancreas, after marking their secretory granules with gold particles. Pancreatic islets were isolated from adult male Wistar rats by the intraductal perfusion of collagenase technique. The islets were cultured in RPMI-1640 medium for 3 days and processed for preparation of ultrathin sections. The sections were stained with the indirect immunogold technique for insulin, glucagon, somatostatin, and pancreatic polypeptide. Ultrastructural examination of the cultured islets clearly identified the presence of B, A, D and PP-cells, as indicated by the numerous gold particles concentrated predominantly over the secretory granules. The secretory granules of the various cell types of the cultured islets demonstrated several similarities as well as differences from the recorded results of the corresponding secretory granules of the intact islets. The differences probably reflect a deviation in the underlying mechanisms of synthesis, maturation and secretion of the different secretory products of the cells in the cultured islets as they adapt to the in vitro environment.
Keywords
culture; electron microscope; immunogold; islets of langerhans; pancreas; rat; ultrastructure
Title
Ultrastructural immunogold study on the various cell types of cultured pancreatic islets of adult rats
Journal
Folia Morphologica
Issue
Vol 59, No 4 (2000)
Article type
Original article
Pages
253-262
Published online
2000-08-02
Page views
410
Article views/downloads
1166
Bibliographic record
Folia Morphol 2000;59(4):253-262.
Keywords
culture
electron microscope
immunogold
islets of langerhans
pancreas
rat
ultrastructure
Authors
Mostafa M El-Naggar