title: A novel method for the assessment of cellular composition and beta-cell viability in human islet preparations.
abstract: 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 beta-cell viability. For these reasons we have developed novel methods for the specific assessment of beta-cell content and viability in human islets based on cellular composition analysis through laser scanning cytometry (LSC) coupled with identification of beta-cell-specific apoptosis at the mitochondrial level. Our novel analytical methods hold promise to prospectively analyze clinical islet transplantation preparations and predict functional performance, as suggested by the observed correlation with in vivo analysis of islet potency in immunodeficient rodents.
authors.affiliation: Diabetes Research Institute, University of Miami, Leonard M. Miller School of Medicine, Miami, FL, USA.
authors.forename: Hirohito
authors.initials: H
authors.lastname: Ichii
authors.forename: Luca
authors.initials: L
authors.lastname: Inverardi
authors.forename: Antonello
authors.initials: A
authors.lastname: Pileggi
authors.forename: R Damaris
authors.initials: RD
authors.lastname: Molano
authors.forename: Over
authors.initials: O
authors.lastname: Cabrera
authors.forename: Alejandro
authors.initials: A
authors.lastname: Caicedo
authors.forename: Shari
authors.initials: S
authors.lastname: Messinger
authors.forename: Yoshikazu
authors.initials: Y
authors.lastname: Kuroda
authors.forename: Per-Olof
authors.initials: PO
authors.lastname: Berggren
authors.forename: Camillo
authors.initials: C
authors.lastname: Ricordi
mesh_terms: D017209:Apoptosis; D002470:Cell Survival / Q000187:drug effects; D016207:Cytokines / Q000494:pharmacology; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007515:Islets of Langerhans / Q000502:physiology*; D047569:Laser Scanning Cytometry; D008928:Mitochondria / Q000502:physiology; D009569:Nitric Oxide / Q000494:pharmacology; D009942:Organometallic Compounds; D016877:Oxidants / Q000494:pharmacology; D013194:Staining and Labeling; D019737:Transplants
chemical_list: D016207:Cytokines; D009942:Organometallic Compounds; D016877:Oxidants; C110932:tetramethyl rhodamine ethyl ester; D009569:Nitric Oxide; D006861:Hydrogen Peroxide