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


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-alpha) inducible vascular cell adhesion molecule-1. IFNG (previously known as IFN-gamma) 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
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.
Favaro
E
E
Department of Internal Medicine and Center of Experimental Medicine (CeRMS), University of Turin, Turin, Italy.
Bottelli
A
A
Lozanoska-Ochser
B
B
Ferioli
E
E
Huang
G C
GC
Klein
N
N
Chiaravalli
A
A
Perin
P Cavallo
PC
Camussi
G
G
Peakman
M
M
Conaldi
P G
PG
Zanone
M M
MM
D002448:Cell Adhesion; D002460:Cell Line; D002465:Cell Movement; D049109:Cell Proliferation; D002478:Cells, Cultured; D003922:Diabetes Mellitus, Type 1 / Q000503:physiopathology; D003924:Diabetes Mellitus, Type 2 / Q000503:physiopathology; D019040:E-Selectin / Q000032:analysis; D042783:Endothelial Cells / Q000737:chemistry / Q000166:cytology* / Q000276:immunology*; D005434:Flow Cytometry; D006684:HLA-DR Antigens / Q000032:analysis; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000032:analysis; D007515:Islets of Langerhans / Q000098:blood supply* / Q000737:chemistry / Q000166:cytology / Q000276:immunology*; D007963:Leukocytes, Mononuclear / Q000166:cytology; D008855:Microscopy, Electron, Scanning; D008856:Microscopy, Fluorescence; D010641:Phenotype; D013539:Simian virus 40; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis
D019040:E-Selectin; D006684:HLA-DR Antigens; D014409:Tumor Necrosis Factor-alpha; D018799:Intercellular Adhesion Molecule-1