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

BMC Veterinary Research

Hans J Nauwynck¹  Hannah L Dewerchin¹  Annelike Dedeurwaerder¹  Lowiese MB Desmarets¹  Dominique AJ Olyslaegers¹ 

[1] Department of Virology, Parasitology and Immunology, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, B-9820 Merelbeke, Belgium

关键词: Vena cava;    Aorta;    hTERT;    SV40LT;    Immortalization;    Feline;    Endothelial cells;   

Others  :  1119468
DOI  :  10.1186/1746-6148-9-170

 received in 2013-06-13, accepted in 2013-08-16,  发布年份 2013

【 摘 要 】

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.

【 授权许可】

   
2013 Olyslaegers et al.; licensee BioMed Central Ltd.

【 图 表 】

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