Article

Diabetologia

, Volume 48, Issue 12, pp 2552-2562

Primary and immortalised human pancreatic islet endothelial cells: phenotypic and immunological characterisation

  • E. FavaroAffiliated withDepartment of Internal Medicine and Center of Experimental Medicine (CeRMS), University of Turin
  • , A. BottelliAffiliated withDepartments of Medicine and Public Health and Pathology, University of Insubria
  • , B. Lozanoska-OchserAffiliated withDepartments of Immunobiology and Diabetes and Internal Medicine, Guy's, King's and St. Thomas's School of Medicine
  • , E. FerioliAffiliated withDepartments of Medicine and Public Health and Pathology, University of Insubria
  • , G. C. HuangAffiliated withDepartments of Immunobiology and Diabetes and Internal Medicine, Guy's, King's and St. Thomas's School of Medicine
  • , N. KleinAffiliated withDepartment of Immunobiology, Institute of Child Health
  • , A. ChiaravalliAffiliated withDepartments of Medicine and Public Health and Pathology, University of Insubria
  • , P. Cavallo PerinAffiliated withDepartment of Internal Medicine and Center of Experimental Medicine (CeRMS), University of Turin
  • , G. CamussiAffiliated withDepartment of Internal Medicine and Center of Experimental Medicine (CeRMS), University of Turin
    • , M. PeakmanAffiliated withDepartments of Immunobiology and Diabetes and Internal Medicine, Guy's, King's and St. Thomas's School of Medicine
    • , P. G. ConaldiAffiliated withDepartments of Medicine and Public Health and Pathology, University of Insubria
    • , M. M. ZanoneAffiliated withDepartment of Internal Medicine and Center of Experimental Medicine (CeRMS), University of TurinDipartimento di Medicina Interna Email author 

Abstract

Aims/hypothesis

Studies on the biology of the microvascular endothelial cells (MECs) that surround and penetrate the pancreatic islets are hampered by difficulties in isolating and culturing large numbers of pure cells. We aimed to morphologically and functionally characterise primary MECs purified and cultured from human islets, and to establish a simian virus 40 (SV40)-immortalised cell line from these primary cultures.

Materials and methods

Human islet MECs were extracted and purified using anti-CD105 coated immunomagnetic beads, and endothelial markers and surface molecules analysed by flow cytometric analysis. An immortalised cell line was then established by using a chimeric adeno5/SV40 virus.

Results

Islet MECs expressed classic and specific endothelial markers, a high basal level of intercellular adhesion molecule-1, and low levels of E-selectin and TNF (previously known as TNF-α) inducible vascular cell adhesion molecule-1. IFNG (previously known as IFN-γ) induced expression of HLA class II molecules. The immortalised islet MECs expanded rapidly, exhibited increased DNA synthesis, and were passaged approximately 30 times, without signs of senescence. They retained the endothelial characteristics of the parental cells, and behaved as the primary cells in terms of TNF stimulation of expression of adhesion molecules and support of leucocyte adhesion and transmigration.

Conclusions/interpretation

The immortalised islet MECs that we have established could effectively represent a substitute for primary counterparts for in vitro studies on the role of the microvasculature in pathophysiological processes involved in type 1 and type 2 diabetes.

Keywords

Adhesion molecules Endothelial cells Islets of Langerhans Large T antigen SV40