A Novel Method for the Assessment of Cellular Composition and Beta‐Cell Viability in Human Islet Preparations

@article{Ichii2005ANM,
  title={A Novel Method for the Assessment of Cellular Composition and Beta‐Cell Viability in Human Islet Preparations},
  author={Hirohito Ichii and Luca A Inverardi and Antonello Pileggi and Ruth Damaris Molano and Over Cabrera and Alejandro Caicedo and Shari Messinger and Yoshikazu Kuroda and P O Berggren and Camillo Ricordi},
  journal={American Journal of Transplantation},
  year={2005},
  volume={5}
}
Current methodologies to evaluate islet cell viability are largely based on tests that assess the exclusion of DNA‐binding dyes. While these tests identify cells that have lost selective membrane permeability, they do not allow us to recognize apoptotic cells, which do not yet stain with DNA‐binding dyes. Furthermore, current methods of analysis do not discriminate between cell subsets in the preparation and, in particular, they do not allow for selectively defining β‐cell viability. 
Assessment of Beta Cell Viability
TLDR
This unit contains detailed protocols for the simultaneous identification of the human pancreatic β cells and determination of their viability by flow cytometry and the method for assessing the mitochondrial functionality byflow cytometry, simple to perform and sufficient to detect the viability of β cells in human islet preparations.
Comparison of two techniques for assessing pancreatic islet viability: flow cytometry and fluorescein diacetate/propidium iodide staining
TLDR
The aim of this study was to compare islet viability as assessed by flow cytometry and the FDA/PI assay and to suggest a quantitative method may be useful.
Islet assessment for transplantation
TLDR
More sensitive and reliable islet viability and potency tests have been recently developed and tested and those evaluating mitochondrial function are most promising, correlate with transplant outcomes in mice, and are currently being evaluated in the clinical setting.
Determination of Islet Viability Using a Zinc-Specific Fluorescent Dye and a Semiautomated Assessment Method
TLDR
NG-DCF/PI staining is easy and reliable, and this method permits highly objective islet-specific viability assessments, and a semiautomated measurement was introduced to determine objective and reproducible results.
Quantitative Assessment of &bgr;-Cell Apoptosis and Cell Composition of Isolated, Undisrupted Human Islets by Laser Scanning Cytometry
TLDR
The analysis of whole, nondissociated islets for cell composition and &bgr;-cell apoptosis using laser scanning cytometry gives reliable and reproducible results and could be performed both before islet transplantation and on preserved cell blocks at any time in future.
Quantitative analysis of cell composition and purity of human pancreatic islet preparations
TLDR
Both EM and LM provide accurate methods to determine the cell composition of human islet preparations and can help us understand many of the discrepancies of islet composition in the literature.
beta-Cell specific cytoprotection by prolactin on human islets.
Characterization of pancreatic ductal cells in human islet preparations
TLDR
The precise characterization and assessment of these cells in addition to β-cells in human islet cell products may be of assistance in understanding their contribution to islet engraftment and in developing strategies to enhance islet graft function.
Current specifications for lot release prior to clinical islet transplantation
TLDR
Assays evaluating mitochondrial function, specifically mitochondrial membrane potential, bioenergetic status, and cellular oxygen consumption rate, especially when conducted with intact islets, appear most promising in evaluating their quality prior to ICT.
A Simplified Approach to Human Islet Quality Assessment
TLDR
This study demonstrates that a multiparametric approach using objective assessments focused on islet cell mitochondrial integrity and in vitro function can provide data predictive of in vivo function.
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