Islet yield after different methods of pancreatic Liberase delivery.
OBJECTIVE
Enzymatic digestion of the pancreas is a fundamental step in islet isolation and there are many ways to administer the enzyme during procurement. The aim of this study was to evaluate the influence of different methods of Liberase delivery during pancreas harvest on the quality and quantity of islets.


METHODS
Depending on the type of Liberase delivery, 4 groups were created. Group 1 was intraductal, Group 2 was interstitial, Group 3 was intragallbladder, and Group 4 was no infusion of enzyme. After injection, the pancreata were harvested, digested in Liberase solution, mechanically disrupted, and purified using discontinuous gradient centrifugation. After 24-hour culture, the number, purity, and viability of the isolated islets were determined.


RESULTS
Intraductal injection of the enzyme yielded statistically significantly more islets per mouse when compared with interstitial, intragallbladder, and no injection administration. Although there was a trend toward better islet purity and viability for Group 1, this was not statistically significant.


CONCLUSIONS
Intraductal administration is the best enzyme delivery method for pancreatic islet isolation. The pancreatic ducts are the most anatomic and physiological way to transport the enzyme uniformly inside the pancreas, determining an adequate digestion and better islet quantity and quality when compared with other delivery methods.
Andrades
P
P
Division of Transplant Immunology, Department of Surgery, University of Alabama in Birmingham, Birmingham, AL 35294-0019, USA. patricio@uab.edu
Asiedu
C
C
Ray
P
P
Rodriguez
C
C
Goodwin
J
J
McCarn
J
J
Thomas
J M
JM
D000818:Animals; D002469:Cell Separation / Q000379:methods; D017364:Collagenases / Q000008:administration & dosage; D007515:Islets of Langerhans / Q000166:cytology*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D010179:Pancreas / Q000166:cytology*; D013820:Thermolysin / Q000008:administration & dosage; D020858:Tissue and Organ Harvesting / Q000379:methods
D017364:Collagenases; C106635:Liberase; D013820:Thermolysin