Document Detail


Quantitative assessment of β-cell apoptosis and cell composition of isolated, undisrupted human islets by laser scanning cytometry.
MedLine Citation:
PMID:  20697327     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND: Assays for assessing human islet cell quality, which provide results before transplantation, would be beneficial to improve the outcomes for islet transplantation therapy. Parameters such as percent β-cell apoptosis and cell composition are found to vary markedly between different islet preparations and may serve as markers of islet quality. We have developed fluorescence-based assays using laser scanning cytometry for assessing β-cell apoptosis and islet cell composition on serial sections of intact isolated islets.
METHODS: Isolated human islets were fixed in formalin and embedded in paraffin. Serial sections were immunostained for the pancreatic hormones and acinar and ductal cell markers. DNA fragmentation was used to label apoptotic cells. Stained cells were quantified using an iCys laser scanning cytometer.
RESULTS: Islet preparations from 102 human pancreatic islet isolations were analyzed. For the whole set of islet preparations, we found a mean islet cell composition of 54.5%±1.2% insulin-positive, 33.9%±1.2% glucagon, 12.1%±0.7% somatostatin, and 1.5%±0.2% pancreatic polypeptide-positive cells. The apoptotic β cells were 2.85%±0.4% with a range of 0.27% to 18.3%. The percentage of apoptotic β cells correlated well (P<0.0001, n=59) with results obtained in vivo by transplantation of the corresponding islets in diabetic NODscid mice.
CONCLUSIONS: The analysis of whole, nondissociated islets for cell composition and β-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. Thus, they can be a powerful tool for islet quality assessment.
Authors:
Ivan Todorov; Indu Nair; Alina Avakian-Mansoorian; Jeffrey Rawson; Keiko Omori; Taihei Ito; Luis Valiente; Itzia Iglesias-Meza; Chris Orr; Keh-Dong Shiang; Kevin Ferreri; Ismail H Al-Abdullah; Yoko Mullen; Fouad Kandeel
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural    
Journal Detail:
Title:  Transplantation     Volume:  90     ISSN:  1534-6080     ISO Abbreviation:  Transplantation     Publication Date:  2010 Oct 
Date Detail:
Created Date:  2010-10-21     Completed Date:  2010-11-16     Revised Date:  2013-05-29    
Medline Journal Info:
Nlm Unique ID:  0132144     Medline TA:  Transplantation     Country:  United States    
Other Details:
Languages:  eng     Pagination:  836-42     Citation Subset:  IM    
Affiliation:
Department of Diabetes and Metabolic Diseases Research, Beckman Research Institute of the City of Hope, Duarte, CA, USA. itodorov@coh.org
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MeSH Terms
Descriptor/Qualifier:
Apoptosis
Glucagon / secretion
Humans
Insulin / secretion
Insulin-Secreting Cells / secretion*
Islets of Langerhans / cytology,  physiology*
Pancreatic Polypeptide / secretion
Somatostatin / secretion
Grant Support
ID/Acronym/Agency:
U42 RR16607/RR/NCRR NIH HHS
Chemical
Reg. No./Substance:
0/Insulin; 51110-01-1/Somatostatin; 59763-91-6/Pancreatic Polypeptide; 9007-92-5/Glucagon
Comments/Corrections

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