The need for analytical methods that are predictive of the quality and potency of cellular products has been recognized. Herein we describe the protocol for an objective quantification of cellular composition and fractional β-cell viability in pancreatic islet cell products. The method is based on the use of Laser Scanning Cytometry (LSC) and Fluorescence-Activated Cell Sorting (FACS) techniques performed on dissociated islet cells. Analysis of human islet preparations with these techniques allows for the definition of β-cell mass and viability, and could provide valuable potency assessment of human islets prior to transplantation.
This work was supported in part by National Institute of Health - NCRR grants for General Clinical Research Center MO1RR16587, NIDDK 1R01-DK55347-IU42RR016603, 5U01DK70460-02, 5R01DK025802-23, Islet Cell Resources grant 5U42RR016603, and Clinical Islet transplant Consortium grants U01DK070460 and 1UL1TR000460; the Juvenile Diabetes Research Foundation International grants 4-200-946 and 4-2004-361; and the Diabetes Research Institute Foundation (www.DiabetesResearch.org). HI was partially supported by a fellowship from the Japanese Society for the promotion of science. The authors are grateful to the members of the Human Cell Processing Facility, of the Preclinical Cell Processing and Translational Models Core, the Cell Transplant Center, of the Clinical Islet Transplant Program, General Clinical Research Center, of the Imaging Core at the Diabetes Research Institute, and of Administrative Offices at the Diabetes Research Institute and of the Organ Procurement Organizations for the continuous enthusiasm and support to our program.
To cite this article
Cellular Composition and Fractional β-Cell Viability Assay Protocol at the Diabetes Research Institute – University of Miami
CellR4 2014; 2 (4): e1121
Published online: 28 Jul 2014