Assessment of human islet isolation with four different collagenases.

Abstract

BACKGROUND:

The isolation of islets from the human pancreas critically depends on the efficiency of the digestive enzymes. Liberase HI had been used as a standard preparation until the issues concerning bovine spongiform encephalopathy. Thus, we must now use other collagenases for clinical islet transplantation, four of which we have evaluated herein.

METHODS:

The digestion of each of 17 pancreata from brain-dead donors was performed using the following collagenases: Liberase HI (HI; Roche, n = 9); Liberase MTF C/T (MTF; Roche, n = 4); Collagenase NB1 Premium Grade (NB1; Serva, n = 7); or Clzyme Collagenase HA (CI, VitaCyte, n = 4). Islet isolations were based on the Edmonton protocol for HI, whereas our modified islet isolation method was used for the three new enzymes (MTF, NB1, and CI).

RESULTS:

There were no significant differences in donor age, body mass index, pancreas size, and cold ischemic time among the four groups. The phase I time in the NB1 group was significantly shorter than in the CI group (P = .0014). The prepurification IEQ/g in the HI group was significantly lower than the others (P = .0003 vs MTF, .0007 vs NB1, and .0009 vs CI, respectively). The postpurification IEQ/g in the MTF group was significantly higher than in the HI group (P = .006). The viability in the NB1 group was significantly greater than the HI group (P = .003).

CONCLUSION:

Three new enzymes (MTF, NB1, and CI) may enable us to obtain higher islet yields than with HI.

• Authors: M Shimoda, H Noguchi, B Naziruddin, Y Fujita, D Chujo, M Takita, H Peng, Y Tamura, G S Olsen, K Sugimoto, T Itoh, N Onaca, M F Levy, P A Grayburn, S Matsumoto
• Journal: Transplantation proceedings (Transplant Proc) 2010 Jul-Aug Vol. 42 Issue 6 Pg. 2049-51 ISSN: 1873-2623 [Electronic] United States
• PMID: 20692404 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright 2010 Elsevier Inc. All rights reserved.

Chemical References

• Collagenases
• Liberase
• Thermolysin

Topics

• Adult
• Animals
• Body Mass Index
• Brain Death
• Cattle
• Cell Separation (methods)
• Collagenases (metabolism)
• Encephalopathy, Bovine Spongiform (pathology)
• Humans
• Islets of Langerhans (cytology, pathology)
• Middle Aged
• Organ Size
• Pancreas (anatomy & histology, pathology)
• Thermolysin (metabolism)
• Tissue Donors