Isolation of Human Pancreatic Islets from the Indian Population - Unveiling the Synergistic Interplay between different blends of Collagenases and Neutral Proteases

Abstract

Innovative and effective therapeutic approaches for diabetes such as islet cell transplantation are much needed as diabetes increases the risk of heart failure. In India, the success rate of human clinical islet transplantation (allo-or auto-) mandates sufficiency of functional islets from the pancreas of either brain-dead or chronic pancreatitis donors. Human islet transplantation outcome is significantly affected by collagenases, which decides the yield and quality of isolated pancreatic islet cells. Comparison of collagenase blends carried out in a few islet transplantation centers yields conflicting results for islet quantity and quality. Therefore, we have researched on the right blend of enzymes to achieve an efficient pancreatic islet release, first-ever in Indian population. Our prospective approach evaluated the outcome of islet isolations performed in humans with three different collagenases, namely Serva Collagenase NB1, Roche Liberase and Vitacyte supplemented with Neutral Protease NB. Eight brain-dead humans served as organ donors for pancreatic islet isolation. We modified the islet isolation method to test these three new enzymes using the traditional split pancreas methodology. The islet isolation outcomes were compared for morphological differences, yield, and viability. This was correlated statistically with the donor characteristics using graph-pad prism analysis. Islet yield was substantial in all the three enzyme blends. But post-purification of islets in the Indian population, we found that the yield was better in Serva collagenase NB1 compared to Vitacyte and Roche liberase. For every gram of pancreas, the islet yield, proportion and morphology of islet cells (both free and intact) were higher in Serva collagenase compared to the other two enzyme blends.