open access

Vol 59, No 4 (2000)
Original article
Submitted: 2012-02-06
Published online: 2000-08-02
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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.

open access

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.
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Keywords

culture; electron microscope; immunogold; islets of langerhans; pancreas; rat; ultrastructure

About this article
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

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