Isolation and characterization of immortalized porcine aortic endothelial cell lines.
Primary porcine endothelial cells have a limited life span in culture. After four to five passages, they tend to de-differentiate and eventually reach senescence. The aim of this work was to establish immortalized porcine aortic endothelial cell lines (AOCs) to facilitate in vitro studies of different pathological process involving the endothelium. Primary porcine aortic endothelial cells (PAECs) were transfected with a plasmid containing the SV40 genome and selected on the basis of morphological and phenotypical features. Flow cytometry analysis demonstrated uptake of acetylated low density lipoproteins (Ac-LDL) and constitutive expression of SLA class I, CD29, CD31, CD41/61, CD80/86, CD46, SWC3, and LAMP-1 antigens by all analyzed lines and showed little differences to primary cells. The functional similarity between primary and immortalized endothelial cells was demonstrated in a cytotoxicity assay using a human natural killer cell line (NKL) as effector. The AOCs cell lines should be valuable tools for in vitro study of the human immune response against pig endothelial cells. In addition, they would be very useful to gain insight in the pathogenesis of some viral haemorrhagic diseases of pig such as African swine fever (ASF) or classical swine fever (CSF).
Unidad de Trasplante, Departamento de Cirugía, Hospital Universitario Virgen de la Arrixaca, El Palmar, 30120 Murcia, Spain.
Ana
A
Carrillo
Sonia
S
Chamorro
Manuel
M
Rodríguez-Gago
Belén
B
Alvarez
María Jesús
MJ
Molina
José Ignacio
JI
Rodríguez-Barbosa
Alicia
A
Sánchez
Pablo
P
Ramírez
Antonio
A
Muñoz
Javier
J
Domínguez
Pascual
P
Parrilla
José
J
Yélamos
D000818:Animals; D001011:Aorta / Q000166:cytology / Q000276:immunology; D018929:Cell Culture Techniques; D002461:Cell Line, Transformed; D003601:Cytotoxicity Tests, Immunologic / Q000662:veterinary; D004730:Endothelium, Vascular / Q000166:cytology* / Q000276:immunology; D005434:Flow Cytometry / Q000662:veterinary; D006801:Humans; D007694:Killer Cells, Natural / Q000276:immunology; D008077:Lipoproteins, LDL / Q000276:immunology / Q000493:pharmacokinetics; D013539:Simian virus 40 / Q000235:genetics; D013552:Swine; D014162:Transfection
D008077:Lipoproteins, LDL; C032629:acetyl-LDL