The effect of UW solution and its components on the collagenase digestion of human and porcine pancreas.
University of Wisconsin (UW) solution is used extensively as a cold storage solution during the procurement and transport of the pancreas prior to islet isolation. However, it has been observed that UW inhibits the collagenase digestion phase of human but not porcine islet isolation, resulting in poor islet yields and islets of poor viability. The aim of this study was, therefore, to confirm this species difference and to determine which components of UW are responsible for the inhibition in the human. In the initial experiment, blocks of human and porcine pancreas (n = 7) were incubated in test tubes containing collagenase at a concentration of 4 mg/mL at 37 degrees C dissolved in 4 mL of either Hanks' solution or UW. Every 5 min the tubes were manually shaken and the degree of tissue dissociation scored on a scale of + and 3+. Our results confirm the inhibition of collagenase digestion in the human but not the pig. Using the same methodology, we then investigated the components of UW that were causing the observed inhibition in the human pancreas (n = 7). This time the collagenase was dissolved in individual or combinations of UW components. Using Hank's as a control, the results were then expressed as a median ratio. The components found to be most inhibitory were magnesium, the Na+/K+ ratio, hydroxyethyl starch (HES), and adenosine. Allopurinol in combination with either lactobionate or glutathione was markedly inhibitory (i.e., median ratio 1.8 and 1.9, respectively). The most inhibitory solution tested was combination of the three components raffinose, glutathione, and lactobionate (median ratio 2.1). This combination was almost as inhibitory as UW itself (median ratio 2.7). These findings are essential for the development of effective cold-storage solutions for the human pancreas that do not inhibit the subsequent collagenase digestion phase of islet isolation.
Contractor
H H
HH
Department of Surgery, University of Leicester, Royal Infirmatory, UK.
Johnson
P R
PR
Chadwick
D R
DR
Robertson
G S
GS
London
N J
NJ
D000241:Adenosine / Q000494:pharmacology; D000328:Adult; D000493:Allopurinol / Q000494:pharmacology; D000818:Animals; D017364:Collagenases / Q000378:metabolism*; D003470:Culture Media; D005978:Glutathione / Q000494:pharmacology; D006801:Humans; D006895:Hydroxyethyl Starch Derivatives / Q000494:pharmacology; D007328:Insulin / Q000494:pharmacology; D007515:Islets of Langerhans / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D016381:Islets of Langerhans Transplantation; D008274:Magnesium / Q000494:pharmacology; D008875:Middle Aged; D019188:Organ Preservation Solutions; D010179:Pancreas / Q000201:enzymology* / Q000648:ultrastructure; D011188:Potassium / Q000494:pharmacology; D011887:Raffinose / Q000494:pharmacology; D012964:Sodium / Q000494:pharmacology; D013552:Swine; D014021:Tissue Preservation
D003470:Culture Media; D006895:Hydroxyethyl Starch Derivatives; D007328:Insulin; D019188:Organ Preservation Solutions; C055206:University of Wisconsin-lactobionate solution; D000493:Allopurinol; D012964:Sodium; D017364:Collagenases; D005978:Glutathione; D008274:Magnesium; D000241:Adenosine; D011887:Raffinose; D011188:Potassium