Advancing Diabetes Research: A Novel Islet Isolation Method from Living Donors

Author:

Di Piazza Eleonora1ORCID,Todi Laura1ORCID,Di Giuseppe Gianfranco12ORCID,Soldovieri Laura12ORCID,Ciccarelli Gea12ORCID,Brunetti Michela12ORCID,Quero Giuseppe234,Alfieri Sergio234,Tondolo Vincenzo4,Pontecorvi Alfredo12,Gasbarrini Antonio25ORCID,Nista Enrico Celestino25,Giaccari Andrea12ORCID,Pani Giovambattista2ORCID,Mezza Teresa25ORCID

Affiliation:

1. Endocrinology and Diabetology Unit, Fondazione Policlinico Universitario Gemelli IRCCS, 00168 Roma, Italy

2. Department of Medicine and Translational Surgery, General Pathology Section, Università Cattolica del Sacro Cuore, 00168 Roma, Italy

3. Digestive Surgery Unit, Fondazione Policlinico Universitario Gemelli IRCCS, 00168 Roma, Italy

4. Digestive Surgery Unit, Ospedale Isola Tiberina—Gemelli Isola, 00186 Roma, Italy

5. Pancreas Unit, CEMAD Centro Malattie dell’Apparato Digerente, Medicina Interna e Gastroenterologia, Fondazione Policlinico Universitario Gemelli IRCCS, 00168 Roma, Italy

Abstract

Pancreatic islet isolation is critical for type 2 diabetes research. Although -omics approaches have shed light on islet molecular profiles, inconsistencies persist; on the other hand, functional studies are essential, but they require reliable and standardized isolation methods. Here, we propose a simplified protocol applied to very small-sized samples collected from partially pancreatectomized living donors. Islet isolation was performed by digesting tissue specimens collected during surgery within a collagenase P solution, followed by a Lympholyte density gradient separation; finally, functional assays and staining with dithizone were carried out. Isolated pancreatic islets exhibited functional responses to glucose and arginine stimulation mirroring donors’ metabolic profiles, with insulin secretion significantly decreasing in diabetic islets compared to non-diabetic islets; conversely, proinsulin secretion showed an increasing trend from non-diabetic to diabetic islets. This novel islet isolation method from living patients undergoing partial pancreatectomy offers a valuable opportunity for targeted study of islet physiology, with the primary advantage of being time-effective and successfully preserving islet viability and functionality. It enables the generation of islet preparations that closely reflect donors’ clinical profiles, simplifying the isolation process and eliminating the need for a Ricordi chamber. Thus, this method holds promises for advancing our understanding of diabetes and for new personalized pharmacological approaches.

Funder

Università Cattolica del Sacro Cuore

Italian Ministry of Education, University, and Research

European Foundation for the Study of Diabetes award sponsored by Astra Zeneca and MIUR

Publisher

MDPI AG

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