24 Jan - 26 Jan '16

6th EPITA Winter Symposium & 35th AIDPIT Workshop

The EPITA Board is happy to announce the 6th EPITA Winter Symposium & 35th AIDPIT Workshop that will take place in Innsbruck from the 24th to the 26th of

Poster

Programme Reference:
PP10
Presenting Author:
Heide Brandhorst 1
Oxford, United Kingdom
Category:
Islet Clinical
Title:
COLLAGENASE AF-1: THE FIRST GMP-GRADED ANIMAL-FREE ENZYME FOR HUMAN ISLET ISOLATION
Body:
Aim GMP-graded animal-free (AF) SERVA Collagenase AF-1 and Neutral Protease (NP) AF have recently become available for human islet isolation. This report describes the initial experiences of 3 different islet transplant centres. Methods Human pancreases retrieved with appropriate consent and ethical approval from 34 organ donors (age: 51.4±1.9 years, male:female: 18:16, BMI: 25.3±0.7, cold ischaemia: 7.25±0.32 hours, trimmed pancreas weight: 102.7±5.0 gram) were digested using one vial of 6 different lots of Collagenase AF-1 (2000 – 2583 PZ-U/vial) supplemented with 4 different lots of NP AF in a range of 50 – 160 DMC-U per pancreas. Isolation, culture and quality assessment were performed using standard techniques as previously described. All data are presented as mean ± SEM. Results The variability of pancreas weight (47 – 164 gram) was associated with a wide range of collagenase and NP activities, ranging from 12.7 – 46.6 PZ-U/g (26.0±1.5 PZ-U/g) and 0.4 – 3.0 DMC-U/g (1.5±0.1 U/g), respectively. Recirculation time required for islet release was 21.5±0.9 min (13 – 41 min). Undigested tissue after digestion ranged from 1 – 32 gram corresponding to a percentage of digested tissue of 85.9±1.3%. Post-purification islet yield was 296,494±33,620 IE (27,826 – 823,125 IE) equivalent to 3274±450 IE/g (218 – 10,624 IE/g) with a purity of 55.9±3.2% (21 – 86%). Quality assessment performed after 2 – 4 days of culture demonstrated viability of 88.1±1.5% (75 – 95%) and insulin content per islet of 10.3±2.2 ng (1.6 – 30.4 ng). Insulin release increased from 0.112±0.019 to 0.185±0.027 ng/islet/hrs (P<0.001) after switching from low (1.0 – 2.8 mM) to high (16.7 – 20 mM) glucose concentration, which is equivalent to a stimulation index of 3.7±0.7 (0.7 – 12.7). Transplant rate was 59% of isolations. Conclusions This study demonstrates the efficiency of animal-free SERVA Collagenase AF-1 and Neutral Protease AF for clinical islet isolation and transplantation. The new plant-based production process makes these products a safe new option for the islet field.