The effect of pancreatic warm ischemia on islet isolation in rats and dogs.
Warm ischemia (WI) has been shown to be detrimental to organ function following transplantation. We investigated the effect of increasing warm ischemic time (WIT) on islet isolation in rats and dogs. Rat isolations were performed by collagenase digestion and Ficoll purification after increasing periods of WI. Dog isolations were performed after similarly increasing periods of WI by ductal perfusion with collagenase, counts being performed on unpurified tissue. Viability studies were performed on isolated purified rat islets by in vitro perifusion. Islet counts decreased as WIT increased such that after 45 min WI islet counts were only 45.7% of those at 0 WIT (P less than 0.001) in rats and 52.5% in dogs (P less than 0.002). Islet volumes decreased to 47.0% in rats (P less than 0.001) in rats and 52.5% in dogs (P less than 0.002). period. After 90 min WIT islet counts were down to 15.6% (P less than 0.001) in rats and 23.9% in dogs (P less than 0.001) and volumes were down to 16.0% (P less than 0.001) in rats and 10.9% (P less than 0.001) in dogs. The increased release of insulin in response to dextrose stimulation was abolished after only 30 min WIT as assessed by perifusion. This work suggests that if successful islet isolation is to be performed for clinical transplantation, WI during donor pancreatectomy must be minimized, or techniques must be developed to prevent or reverse the ensuing effects.
Corlett
M P
MP
Department of Surgery, Washington University School of Medicine, St. Louis, Missouri 63110.
Scharp
D W
DW
D000818:Animals; D004285:Dogs; D005947:Glucose / Q000494:pharmacology; D007328:Insulin / Q000096:biosynthesis; D007511:Ischemia / Q000473:pathology*; D007515:Islets of Langerhans / Q000378:metabolism / Q000473:pathology*; D010179:Pancreas / Q000473:pathology*; D010477:Perfusion; D051381:Rats; D011919:Rats, Inbred Strains; D013268:Stimulation, Chemical; D013696:Temperature; D009927:Tissue and Organ Procurement / Q000379:methods
D007328:Insulin; D005947:Glucose