title: Generation and characterization of feline arterial and venous endothelial cell lines for the study of the vascular endothelium.
abstract: BACKGROUND
  The in vitro culture of endothelial cells (ECs) is an indispensable tool for studying the role of the endothelium in physical and pathological conditions. Primary ECs, however, have a restricted proliferative lifespan which hampers their use in long-term studies. The need for standardized experimental conditions to obtain relevant and reproducible results has increased the demand for well-characterized, continuous EC lines that retain the phenotypic and functional characteristics of their non-transformed counterparts.
  
  
  RESULTS
  Primary feline ECs from aorta and vena cava were successfully immortalized through the successive introduction of simian virus 40 large T (SV40LT) antigen and the catalytic subunit of human telomerase (hTERT). In contrast to the parental ECs, the transformed cells were able to proliferate continuously in culture. Established cell lines exhibited several inherent endothelial properties, including typical cobblestone morphology, binding of endothelial cell-specific lectins and internalization of acetylated low-density lipoprotein. In addition, the immortalization did not affect the functional phenotype as demonstrated by their capacity to rapidly form cord-like structures on matrigel and to express cell adhesion molecules following cytokine stimulation.
  
  
  CONCLUSIONS
  The ability to immortalize feline ECs, and the fact that these cells maintain the EC phenotype will enable a greater understanding of fundamental mechanisms of EC biology and endothelial-related diseases. Furthermore, the use of cell lines is an effective implementation of the 3-R principles formulated by Russel and Burch.
authors.affiliation: Department of Virology, Parasitology and Immunology, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, B-9820 Merelbeke, Belgium. hans.nauwynck@ugent.be.
authors.forename: Dominique A J
authors.initials: DA
authors.lastname: Olyslaegers
authors.forename: Lowiese M B
authors.initials: LM
authors.lastname: Desmarets
authors.forename: Annelike
authors.initials: A
authors.lastname: Dedeurwaerder
authors.forename: Hannah L
authors.initials: HL
authors.lastname: Dewerchin
authors.forename: Hans J
authors.initials: HJ
authors.lastname: Nauwynck
mesh_terms: D000818:Animals; D000952:Antigens, Polyomavirus Transforming; D001011:Aorta / Q000166:cytology / Q000502:physiology; D002415:Cats / Q000502:physiology*; D018929:Cell Culture Techniques / Q000662:veterinary*; D002460:Cell Line; D003599:Cytoskeleton / Q000235:genetics / Q000378:metabolism; D042783:Endothelial Cells / Q000502:physiology* / Q000648:ultrastructure; D005786:Gene Expression Regulation; D006801:Humans; D008077:Lipoproteins, LDL / Q000378:metabolism; D037121:Plant Lectins / Q000494:pharmacology; D019098:Telomerase / Q000235:genetics / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism; D014684:Venae Cavae / Q000166:cytology / Q000502:physiology; D014841:von Willebrand Factor
chemical_list: D000952:Antigens, Polyomavirus Transforming; D008077:Lipoproteins, LDL; D037121:Plant Lectins; D014409:Tumor Necrosis Factor-alpha; C036850:Ulex europaeus lectins; C032629:acetyl-LDL; D014841:von Willebrand Factor; C509186:TERT protein, human; D019098:Telomerase