Rat pancreas preparation for cold storage and subsequent islet cell isolation.
Pancreatic islets can be isolated from cold-preserved organs. They contain 60% more beta cells when prepared from organs that were stored for 24 hr in Collins solution supplemented with albumin and benzamidine (CAB) instead of University of Wisconsin (UW) solution. Recovery from CAB-stored organs was similar when CAB was perfused in situ before organ removal or ex vivo after organ harvesting in UW. In situ flush with cold Ringers before ex vivo replacement by CAB resulted in 25% lower recovery of islet beta cells and in higher contamination with nonendocrine and damaged cells. Recovery of beta cells was 50% reduced when cold storage solution was not chased before collagenase digestion of the organ. It is concluded that the isolation of rat islets from cold-preserved organs can be improved by using UW or CAB instead of Ringers for situ perfusion, by cold storage in CAB, and by adequate chase of cold storage solution with physiologic medium before collagenase digestion. These conditions can be tested in current human islet isolation protocols.
Korbutt
G S
GS
Department of Metabolism and Endocrinology, Vrije Universiteit Brussel, Belgium.
Pipeleers
D G
DG
D000241:Adenosine / Q000494:pharmacology; D000493:Allopurinol / Q000494:pharmacology; D000818:Animals; D002469:Cell Separation; D015925:Cryopreservation; D005978:Glutathione / Q000494:pharmacology; D006982:Hypertonic Solutions / Q000494:pharmacology; D007328:Insulin / Q000494:pharmacology; D007515:Islets of Langerhans / Q000166:cytology*; D008297:Male; D009926:Organ Preservation; D019188:Organ Preservation Solutions; D010179:Pancreas; D011887:Raffinose / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar
C024102:Collins' solution; D006982:Hypertonic Solutions; D007328:Insulin; D019188:Organ Preservation Solutions; C055206:University of Wisconsin-lactobionate solution; D000493:Allopurinol; D005978:Glutathione; D000241:Adenosine; D011887:Raffinose