Highly Purified Versus Filtered Crude Collagenase: Comparable Human Islet Isolation Outcomes

This study was designed to retrospectively compare the impact of crude Sigma V collagenase (Sigma V. n = 52) with high-purified Serva NBI collagenase (Serva NBI, n = 42) on human islet isolation outcomes. A three-step filtration was applied to the crude Sigma V to remove endotoxin contamination and impurities; in addition, this process was used as a lot prescreening tool. Isolation outcomes were determined by digestion efficacy, islet yields, purity, viability, glucose-stimulated insulin release, and endotoxin content. The digestion efficacy between Sigma V and Serva NBI was statistically significant (Sigma V: 64.71% vs. Serva NBI: 69.71%, p = 0.0014). However, the islet yields were similar (Sigma V: 23422.58 vs. Serva NBI: 271097 IEq, p = 0.23) between groups. There was no significant purity difference observed in fractions with purities greater than 75%. Viability (Sigma V: 93.3% vs. Serva NBI : 94.8%, p = 0.061) and stimulation indexes (Sigma V: 3.41 vs. Serva NBI: 2.74, p = 0.187) were also similar between the two groups. The impact of cold ischemia and age on the isolation outcome in the Sigma V group was comparable to the Serva NBI group. The endotoxin content of the final products in the filtered Sigma V group was significantly less than that in the high-purified Serva NBI group (0.022 vs. 0.052 EU/ml, p = 0.003). Additionally, in the Sigma V group there was minimal lot to lot variation and no significant loss of enzymatic activity after filtration. These findings indicate that the use of Sigma V or other crude enzyme blends for research pancreata is warranted to reduce isolation costs and increase the amount of islets available for critical islet research. These findings also validate the need for a systematic enzyme analysis to resolve these inconsistencies in overall enzyme quality once and for all.