Experimental islet isolation in porcine pancreas with new enzyme Liberase PI.
The aim of this study was to investigate the results of 20 consecutive porcine islet isolations using a new enzyme Liberase PI. Twenty pancreata were procured for islet isolation, which was performed using modified Ricordi's method with Liberase PI. Quantitation of islet viability staining, insulin stimulation assay, intracellular insulin content/DNA, and in vivo transplantability into diabetic nude mice were examined for quality control. The results were compared between a high-yield group (>2500 IEQ/g pancreas) and a low-yield group (<2500 IEQ/g pancreas). Sufficient amount of purified islets (3000 IEQ/g pancreas) were obtained using the new brand enzyme Liberase PI. These islets showed good quality in structure and functions, which were demonstrated by in vitro and in vivo standard assays. Isolation index (IEQ/number) of the low-yield group was lower than that of high-yield group (0.75 vs 0.86), which means more fragmentation of islets in the low-yield group. There were no differences in function between the two groups. In conclusion, we obtained sufficient numbers of viable, functional islets from porcine pancreas using a new brand enzyme Liberase PI and low-temperature isolation technique. However, overdigestion of islets during the isolation remains to be overcome. Advance in porcine islet isolation technique will in the future make the porcine islet xenotransplantation a reality for the cure of diabetes mellitus.
Kim
S C
SC
Department of Surgery, Ulsan University College of Medicine and Asan Medical Center, Seoul, Korea. drksc@amc.seoul.kr
Han
D J
DJ
Kang
C H
CH
We
Y M
YM
Back
J H
JH
Kim
Y H
YH
Lim
D G
DG
D000818:Animals; D002469:Cell Separation / Q000379:methods; D002470:Cell Survival; D017364:Collagenases / Q000627:therapeutic use*; D007515:Islets of Langerhans / Q000166:cytology*; D023421:Models, Animal; D010179:Pancreas / Q000166:cytology* / Q000201:enzymology; D013552:Swine; D013820:Thermolysin / Q000627:therapeutic use*; D014184:Transplantation, Homologous / Q000379:methods
D017364:Collagenases; C106635:Liberase; D013820:Thermolysin