Immortalized Human Aortic Endothelial Cells

Immortalized human aortic endothelial cells (HAECs) are established by transducing primary HAECs with recombinant lentiviruses carrying the simian virus 40 (SV40) large T antigen or by stably expressing human telomerase reverse transcriptase (hTERT). The advantages of immortalized HAECs are fourfold. First, extended proliferative capacity – whereas primary cells senesce within a few passages, SV40-transduced cells can be cultured beyond 30 passages, with a population doubling time of approximately 34–44 hours, providing a consistent and virtually unlimited cell supply. Second, phenotypic fidelity – immortalized HAECs retain key endothelial characteristics, including cobblestone morphology, expression of endothelial markers such as CD31/PECAM-1, von Willebrand factor (vWF), and VE-cadherin, as well as functional attributes including LDL uptake and formation of tubular structures that mimic natural blood vessel development. They respond to physiological stimuli: proliferation increases upon VEGF stimulation, and treatment with TNFα induces an effective inflammatory response with upregulation of adhesion molecules ICAM-1 and VCAM-1. Third, functional relevance – these cells produce both antithrombotic and thrombotic factors such as tissue plasminogen activator (t-PA) and plasminogen activator inhibitor-1 (PAI-1), and maintain nitric oxide (NO) production. hTERT-immortalized lines exhibit greater eNOS expression and NO activity compared to senescent primary cells. When co-cultured with fibroblasts, they can form neoangiogenic tubular networks responsive to VEGF stimulation and suramin inhibition. Fourth, experimental reproducibility and scalability – the immortalized nature eliminates donor-to-donor variability inherent to primary cultures, enabling consistent, repeatable experiments and high-throughput screening for drug development and cardiovascular disease research. They are amenable to genetic modification including CRISPR-based editing, and have been extensively validated in studies of atherosclerosis, hypertension, thrombosis, angiogenesis, and stent graft compatibility.