Characterization of pancreatic ductal cells in human islet preparations

Hirohito Ichii, Atsushi Miki, Toshiyuki Yamamoto, Ruth Molano, Scott Barker, Atsuyoshi Mita, Rayner Rodriguez Diaz, Dagmar Klein, Ricardo Pastori, Rodolfo Alejandro, Luca A Inverardi, Antonello Pileggi, Camillo Ricordi

Research output: Contribution to journalArticle

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Abstract

Substantial amounts of nonendocrine cells are implanted as part of human islet grafts, and a possible influence of nonendocrine cells on clinical islet transplantation outcome has been postulated. There are currently no product release criteria specific for nonendocrine cells due to lack of available methods. The aims of this study were to develop a method for the evaluation of pancreatic ductal cells (PDCs) for clinical islet transplantation and to characterize them regarding phenotype, viability, and function. We assessed 161 human islet preparations using laser scanning cytometry (LSC/iCys) for phenotypic analysis of nonendocrine cells and flow cytometry (FACS) for PDC viability. PDC and β-cells obtained from different density fractions during the islet cell purification were compared in terms of viability. Furthermore, we examined PDC ability to produce proinflammatory cytokines/chemokines, vascular endothelial growth factor (VEGF) and tissue factor (TF) relevant to islet graft outcome. Phenotypic analysis by LSC/iCys indicated that single staining for CK19 or CA19-9 was not enough for identifying PDCs, and that double staining for amylase and CK19 or CA19-9 allowed for quantitative evaluation of acinar cells and PDC content in human islet preparation. PDC showed a significantly higher viability than β-cells (PDC vs β-cell: 75.5±13.9 and 62.7±18.7%; P<0.0001). Although β-cell viability was independent of its density, that of PDCs was higher as the density from which they were recovered increased. There was no correlation between PDCs and β-cell viability (R2=0.0078). PDCs sorted from high-density fractions produced significantly higher amounts of proinflammatory mediators and VEGF, but not TF. We conclude that PDCs isolated from different fractions had different viability and functions. 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.

Original languageEnglish
Pages (from-to)1167-1177
Number of pages11
JournalLaboratory Investigation
Volume88
Issue number11
DOIs
StatePublished - Nov 1 2008

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Cell Survival
Islets of Langerhans Transplantation
Thromboplastin
Transplants
Islets of Langerhans
Vascular Endothelial Growth Factor A
Laser Scanning Cytometry
Staining and Labeling
Acinar Cells
Amylases
Chemokines
Flow Cytometry
Cytokines
Phenotype

Keywords

  • CA19-9
  • Cytokines
  • Islet transplantation
  • Laser scanning cytometer
  • Pancreatic ductal cells
  • VEGF

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Cell Biology
  • Molecular Biology

Cite this

Characterization of pancreatic ductal cells in human islet preparations. / Ichii, Hirohito; Miki, Atsushi; Yamamoto, Toshiyuki; Molano, Ruth; Barker, Scott; Mita, Atsuyoshi; Rodriguez Diaz, Rayner; Klein, Dagmar; Pastori, Ricardo; Alejandro, Rodolfo; Inverardi, Luca A; Pileggi, Antonello; Ricordi, Camillo.

In: Laboratory Investigation, Vol. 88, No. 11, 01.11.2008, p. 1167-1177.

Research output: Contribution to journalArticle

Ichii, H, Miki, A, Yamamoto, T, Molano, R, Barker, S, Mita, A, Rodriguez Diaz, R, Klein, D, Pastori, R, Alejandro, R, Inverardi, LA, Pileggi, A & Ricordi, C 2008, 'Characterization of pancreatic ductal cells in human islet preparations', Laboratory Investigation, vol. 88, no. 11, pp. 1167-1177. https://doi.org/10.1038/labinvest.2008.87
Ichii H, Miki A, Yamamoto T, Molano R, Barker S, Mita A et al. Characterization of pancreatic ductal cells in human islet preparations. Laboratory Investigation. 2008 Nov 1;88(11):1167-1177. https://doi.org/10.1038/labinvest.2008.87
Ichii, Hirohito ; Miki, Atsushi ; Yamamoto, Toshiyuki ; Molano, Ruth ; Barker, Scott ; Mita, Atsuyoshi ; Rodriguez Diaz, Rayner ; Klein, Dagmar ; Pastori, Ricardo ; Alejandro, Rodolfo ; Inverardi, Luca A ; Pileggi, Antonello ; Ricordi, Camillo. / Characterization of pancreatic ductal cells in human islet preparations. In: Laboratory Investigation. 2008 ; Vol. 88, No. 11. pp. 1167-1177.
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