Culture prior to transplantation preserves the ultrastructural integrity of monkey pancreatic islets.
Transplantation of pancreatic islets represents a promising way of curing type I diabetes (insulin-dependent diabetes mellitus). Culture enables the survival of endocrine tissue awaiting islet transplantation and reduces islet immunogenicity prior to xenografting. In this study, attempts were made to preserve the monkey islets in culture for 7 days and to study the ultrastructure by electron microscopy. The islets were isolated from monkey pancreas by the collagenase digestion method and were separated from acinar cells by dextran density gradient centrifugation. These islets were preserved in a humidified atmosphere of 5% carbon dioxide and 95% air for 7 days. The culture medium used was CMRL-1066. After 7 days of culture the islets were processed for light and electron microscopic studies, which revealed that the cultured islets were intact and maintained their structural integrity. Semi-thin sections of the cultured islets showed morphology with occasional structural alterations at the periphery. Dithizone staining of the cultured islets showed crimson red colour, proving that the islets were pure and without any exocrine contamination. Electron microscopy showed that the cultured islets had well-preserved alpha-, beta- and delta-cells. Different cell types of the monkey pancreatic islets were identified by the presence of their characteristic secretory granules. The ultrastructural characteristics present in hormone-synthesizing cells, i.e. rough-endoplasmic reticulum, Golgi apparatus, mitochondria and secretory granules, were observed as in native islets.
Department of Physiology, Christian Medical College and Hospital, Vellore-632 002, Tamil Nadu, India.
Daisy M
DM
Mythili
S Susama
SS
Patra
Soosaiah
S
Gunasekaran
D000818:Animals; D002478:Cells, Cultured; D007515:Islets of Langerhans / Q000166:cytology / Q000254:growth & development* / Q000648:ultrastructure*; D016381:Islets of Langerhans Transplantation; D008255:Macaca radiata; D008854:Microscopy, Electron; D010179:Pancreas / Q000648:ultrastructure