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