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Filtration is a time-efficient option to Histopaque, providing good-quality islets in mouse islet isolation

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Abstract

Pancreatic islet transplantation is a promising therapy for Type I Diabetes. For many years the method used worldwide for islet purification in both rodent and human islet isolation has been Ficoll-based density gradients, such as Histopaque. However, it is difficult to purify islets in laboratories with staff limitations when large scale isolations are required. We hypothesized that filtration could be a more simple and fast alternative to obtain good quality islets. Four separate islet isolations were performed per method, comparing filtration and Histopaque purification with handpicking as the gold standard method for islet purity. Different parameters of quality were assessed: yield in number of islets per pancreas, purity by dithizone staining, viability by Fluorescein Diacetate/Propidium Iodide vital staining and in vitro functionality assessed by Glucose Stimulated Insulin Secretion. Time efficiency and cost were also analyzed. The overall quality of the islets obtained both by Histopaque and filtration was good. Filtration saved almost 90 % of the time consumed by Histopaque purification, and was also cheaper. However, one-third of the islets were lost. Since human and rodent islets share similar size but different density, filtration appears as a purification method with potential interest in translation to clinic.

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Acknowledgments

M. R. D. conceived the experimental design, researched data, performed experiments, analyzed results and wrote the manuscript. L. C. contributed to discussion and edited the manuscript. The authors acknowledge Dr. Anna Novials (Institut d’Investigacions Biomédiques August Pi i Sunyer, Barcelona, Spain), Dr. Franz Martín (Centro Andaluz de Biología Molecular y Medicina Regenerativa, Sevilla, Spain), Dr. Eduard Montanya (Hospital de Bellvitge-IDIBELL, Barcelona, Spain) and Dr. Simona Marzorati (Ospedale San Raffaele, Milán, Spain) for scientific discussion. We thank Dr. José Ramón Bilbao, Dr. David Fogarty (University of the Basque Country, Leioa, Spain) and Dr. Raquel Malumbres (Centro de Investigación Médica Aplicada, Pamplona, Spain) for editorial assistance. Technical and human support provided by SGIker (UPV/EHU, MICINN, GV/EJ, ERDF and ESF) is gratefully acknowledged. This work was supported by a grant of the Basque Government for Groups of Excellence (IT-472-07) and a Grant of the Department of Industry of the Basque Government (SAIO09-PE09BF01). The authors have no duality of interest to declare.

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Authors and Affiliations

  1. Laboratory of Cell Therapy in Diabetes, Department of Pediatrics, Faculty of Medicine and Odontology, Cruces University Hospital, University of the Basque Country (UPV/EHU), Barrio Sarriena, s/n, 48940, Leioa, Vizcaya, Spain

    Miriam Ramírez-Domínguez & Luis Castaño

  2. Spanish Biomedical Research Centre in Diabetes and Associated Metabolic Disorders (CIBERDEM), Barcelona, Spain

    Miriam Ramírez-Domínguez & Luis Castaño

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  1. Miriam Ramírez-Domínguez
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Correspondence to Miriam Ramírez-Domínguez.

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Ramírez-Domínguez, M., Castaño, L. Filtration is a time-efficient option to Histopaque, providing good-quality islets in mouse islet isolation. Cytotechnology 67, 199–206 (2015). https://doi.org/10.1007/s10616-014-9690-7

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