Human islet isolation using animal-free collagenase and neutral protease
Geraldine Parnaud, Switzerland Click here for Recording
Thursday September 26, 2013 from 15:30 to 16:30
Big Sur Room
Authors and Affilliations

Geraldine Parnaud1, Vanessa Lavallard1, Raphael Meier1, Bedoit Bedat1, Domenico Bosco1, Thierry Berney1

1Cell Isolation and Transplantation Center, Division of Surgical Research, Geneva University Hospital and University of Geneva, Geneva, Switzerland

Presenter

Geraldine Parnaud, Switzerland

Biologist

Surgery

Abstract

BACKGROUND: The quality of the enzymes used to dissociate the pancreas is critical for successful islet isolation. Despite the safety of the enzymes is certified, enzyme production still requires contact with some animal products. To abolish the risk of contamination of animal derived adventitious agents, animal-free collagenase (AF-1) and neutral protease (NP AF) were developed by SERVA. In this study, we assessed the impact of AF-1 and NP AF on islet isolation outcome.

METHODS: We report the results of 10 islet isolations from pancreases digested with SERVA Collagenase NB 1 Premium Grade with Neutral Protease NB High Active Grade (n=5) or with SERVA Collagenase AF-1 with Neutral Protease AF (n=5). We used an enzyme mixture composed of >2000 U of collagenase and 160 U of neutral protease. Islet isolation outcomes and islet quality were determined from the two enzyme preparations and statistically compared using Student’s test analysis.

RESULTS: No significant differences in donor-related factors were found between the groups. There were also no significant difference in time (NB 1 vs. AF-1 : 17.8 ± 4.0 vs. 17.2 ± 1.1 min) and efficacy of digestion (NB1 vs. AF-1: 89 ± 8% vs. 91 ± 2%). Using Collagenase AF-1 islet products encountered release criteria for transplantation from all of the 5 pancreases digested and from 3 of 5 pancreases when using Collagenase NB 1. Islet yields were similar between the two groups with 354110±164470 and 364510±134490 total IEQ/transplantation and 4550±2562 and 4153±3020 IEQ/g pancreas for NB 1 and AF-1 respectively. Furthermore beta-cell secretory function and viability were similar for the two groups.

CONCLUSION:  The enzyme mixture composed of animal free collagenase (AF-1) with animal free neutral protease recovers similar islet yield than the Collagenase NB 1 Premium Grade while retaining islet quality and function. Thus, these enzymes represent a promising and safe new alternative to the enzymes which are currently used for islet isolation.

 

 

 

 

Lectures by Geraldine Parnaud