Primary and immortalised human pancreatic islet endothelial cells: phenotypic and immunological characterisation
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.
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.
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.
- Primary and immortalised human pancreatic islet endothelial cells: phenotypic and immunological characterisation
Volume 48, Issue 12 , pp 2552-2562
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- Adhesion molecules
- Endothelial cells
- Islets of Langerhans
- Large T antigen
- Industry Sectors
- Author Affiliations
- 1. Department of Internal Medicine and Center of Experimental Medicine (CeRMS), University of Turin, Turin, Italy
- 2. Departments of Medicine and Public Health and Pathology, University of Insubria, Varese, Italy
- 3. Departments of Immunobiology and Diabetes and Internal Medicine, Guy's, King's and St. Thomas's School of Medicine, London, UK
- 4. Department of Immunobiology, Institute of Child Health, London, UK
- 5. Dipartimento di Medicina Interna, Corso Dogliotti 14, 10126, Torino, Italy