title: Central necrosis in isolated hypoxic human pancreatic islets: evidence for postisolation ischemia.
abstract: A variety of explanations have been provided to elucidate the requirement of the large islet mass that is essential for a successful treatment of patients with type I diabetes by intrahepatic transplantation. The purpose of this study was to investigate islet cell survival under the effect of prolonged hypoxia and/or nutrient withdrawal, which mimics posttransplantation environment of transplanted islets in the liver. We studied the influence of 24 h of hypoxia (1% O2) in intact isolated human and rat islets as well as the effect of combined oxygen/nutrient deprivation in a mouse insulinoma cell line (MIN6). In intact human islets, 24 h of hypoxia led to central necrosis combined with apoptotic features such as nuclear pyknosis and DNA fragmentation. In the course of hypoxic treatment, ultrastructural analysis demonstrated a gradual transition from an apoptotic to a necrotic morphology particularly pronounced in central areas of large islets. In MIN6 cells, on the other hand, hypoxia led to a twofold (p < 0.01) increase in caspase-3 activity, an indicator of apoptosis, but not to necrosis, as determined by release of lactate dehydrogenase (LDH). Only in combination with nutrient/serum deprivation was a marked increase in LDH release observed (sixfold vs. control, p < 0.01). We therefore conclude that, similar to MIN6 cells, central necrosis in isolated hypoxic islets is the result of the combined effects of hypoxia and nutrient/serum deprivation, most likely due to limited diffusion. Provided that transplanted islets undergo a similar fate as shown in our in vitro study, future emphasis will require the development of strategies that protect the islet graft from early cell death and accelerate the revascularization process.
authors.affiliation: Department of Visceral and Transplant Surgery, University Hospital Zurich, 8091 Zurich, Switzerland.
authors.forename: Mauro
authors.initials: M
authors.lastname: Giuliani
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Moritz
authors.forename: Elvira
authors.initials: E
authors.lastname: Bodmer
authors.forename: Daniel
authors.initials: D
authors.lastname: Dindo
authors.forename: Patrick
authors.initials: P
authors.lastname: Kugelmeier
authors.forename: Roger
authors.initials: R
authors.lastname: Lehmann
authors.forename: Max
authors.initials: M
authors.lastname: Gassmann
authors.forename: Peter
authors.initials: P
authors.lastname: Groscurth
authors.forename: Markus
authors.initials: M
authors.lastname: Weber
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000328:Adult; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D015687:Cell Hypoxia / Q000502:physiology; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D006801:Humans; D007511:Ischemia / Q000503:physiopathology*; D007515:Islets of Langerhans / Q000378:metabolism / Q000473:pathology* / Q000648:ultrastructure; D016381:Islets of Langerhans Transplantation; D008297:Male; D051379:Mice; D046529:Microscopy, Electron, Transmission; D008875:Middle Aged; D009336:Necrosis; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D016895:Culture Media, Serum-Free; D000255:Adenosine Triphosphate; C505407:CASP3 protein, human; C505409:Casp3 protein, mouse; C505410:Casp3 protein, rat; D053148:Caspase 3; D020169:Caspases