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Generation and characterization of feline arterial and venous endothelial cell lines for the study of the vascular endothelium

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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. Conclusion: 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.
Keywords
Feline, Immortalization, SIMIAN-VIRUS-40, CULTURE, BIOLOGY, EXPRESSION, LIFE-SPAN, LARGE T-ANTIGEN, Vena cava, Aorta, SV40LT, hTERT, Endothelial cells, IMMORTALIZATION, TELOMERASE ACTIVITY, HUMAN FIBROBLASTS, IMMUNODEFICIENCY VIRUS

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Chicago
Olyslaegers, Dominique, Lowiese Desmarets, Annelike Dedeurwaerder, Hannah Dewerchin, and Hans Nauwynck. 2013. “Generation and Characterization of Feline Arterial and Venous Endothelial Cell Lines for the Study of the Vascular Endothelium.” Bmc Veterinary Research 9.
APA
Olyslaegers, D., Desmarets, L., Dedeurwaerder, A., Dewerchin, H., & Nauwynck, H. (2013). Generation and characterization of feline arterial and venous endothelial cell lines for the study of the vascular endothelium. BMC VETERINARY RESEARCH, 9.
Vancouver
1.
Olyslaegers D, Desmarets L, Dedeurwaerder A, Dewerchin H, Nauwynck H. Generation and characterization of feline arterial and venous endothelial cell lines for the study of the vascular endothelium. BMC VETERINARY RESEARCH. 2013;9.
MLA
Olyslaegers, Dominique, Lowiese Desmarets, Annelike Dedeurwaerder, et al. “Generation and Characterization of Feline Arterial and Venous Endothelial Cell Lines for the Study of the Vascular Endothelium.” BMC VETERINARY RESEARCH 9 (2013): n. pag. Print.
@article{4157883,
  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.
Conclusion: 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.},
  articleno    = {170},
  author       = {Olyslaegers, Dominique and Desmarets, Lowiese and Dedeurwaerder, Annelike and Dewerchin, Hannah and Nauwynck, Hans},
  issn         = {1746-6148},
  journal      = {BMC VETERINARY RESEARCH},
  keyword      = {Feline,Immortalization,SIMIAN-VIRUS-40,CULTURE,BIOLOGY,EXPRESSION,LIFE-SPAN,LARGE T-ANTIGEN,Vena cava,Aorta,SV40LT,hTERT,Endothelial cells,IMMORTALIZATION,TELOMERASE ACTIVITY,HUMAN FIBROBLASTS,IMMUNODEFICIENCY VIRUS},
  language     = {eng},
  pages        = {11},
  title        = {Generation and characterization of feline arterial and venous endothelial cell lines for the study of the vascular endothelium},
  url          = {http://dx.doi.org/10.1186/1746-6148-9-170},
  volume       = {9},
  year         = {2013},
}

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