Evaluation of collagenase gold plus BP protease in isolating islets from human pancreata

Islets. 2018 Mar 4;10(2):51-59. doi: 10.1080/19382014.2017.1417716. Epub 2018 Feb 2.

Abstract

Selection of enzymes for optimal pancreas digestion is essential for successful human islet isolations. The aim of this study was to evaluate the efficacy and outcome of using Collagenase Gold plus BP protease (VitaCyte) (n = 8) by comparing it to two commercially available enzymes, Liberase MTF C/T (Roche) (n = 48) and Collagenase NB1/NP (Serva) (n = 15). The isolation outcomes were assessed by islet counting, viability, glucose-stimulated oxygen consumption rate (OCR), and successful graft-rate following transplantation in diabetic NOD scid mice. The pancreas donor characteristics were not significantly different between the tested enzyme groups regarding their BMI, pancreas weight, cold ischemia time (CIT) and HbA1c. The results show that digested tissue volume was not statistically significant between the VitaCyte enzyme (34.25 ± 5.4 mL) and the Roche enzyme (55.25 ± 3.42 mL, p = 0.073), however, this was significant with Serva enzyme (64.07 ± 7.95 mL, p = 0.020). Interestingly, the islet yields were not statistically different between all enzyme groups. Moreover, when islets were transplanted into NOD scid mice, the reversal rate of diabetes for the VitaCyte enzyme group was similar to all enzyme groups. In conclusion, the effectiveness of Collagenase Gold plus BP protease is comparable to the MTF C/T and the Collagenase NB1/NP enzymes; the low cost could facilitate the use of more pancreata for islet isolations.

Keywords: Liberase MTF C/T; collagenase NB1; collagenase gold; islet isolation; neutral protease.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Animals
  • Cell Separation / methods*
  • Cell Survival
  • Collagenases*
  • Graft Survival
  • Humans
  • Islets of Langerhans / cytology*
  • Islets of Langerhans Transplantation*
  • Male
  • Mice
  • Mice, Inbred NOD
  • Mice, SCID
  • Middle Aged
  • Oxygen Consumption
  • Peptide Hydrolases*
  • Retrospective Studies
  • Thermolysin
  • Young Adult

Substances

  • Peptide Hydrolases
  • Collagenases
  • Liberase
  • Thermolysin