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Updated: Jul 19, 2026
Human Saphenous Vein Endothelial Cell Isolation and Exposure to Controlled Levels of Shear Stress and Stretch
Published on: April 21, 2023
Phenotype gradation of human saphenous vein endothelial cells from cardiovascular disease subjects
C Krishna Prasad1, K Jayakumar, Lissy K Krishnan
1Thrombosis Research Unit, Biomedical Technology Wing, Sree Chitra Tirunal Institute for Medical Sciences and Technology, Trivandrum, India.
Insights
A novel biomimetic matrix model helps study endothelial cell (EC) function in cardiovascular diseases (CVDs). This model better maintains EC phenotype, crucial for understanding EC dysfunction and developing targeted therapies.
Area of Science:
- Vascular Biology
- Biomaterials Science
- Cardiovascular Research
Background:
- Endothelial cell (EC) phenotype alterations are key in cardiovascular diseases (CVDs), but their relationship with extracellular matrix (ECM) changes is unclear.
- In vitro EC studies are limited by dedifferentiation in subcultures, hindering research into EC dysfunction.
Purpose of the Study:
- To characterize the phenotype of human saphenous vein endothelial cells (HSVECs) from CVD patients using an in vitro biomimetic matrix model.
- To assess the influence of matrix composition on EC phenotype, proliferation, and apoptosis.
Main Methods:
- Cultured HSVECs from CVD patients on a biomimetic matrix and conventional polystyrene.
- Analyzed mRNA expression and synthesis of key molecules: von Willebrand factor (vWF), plasminogen activation inhibitor (PAI), tissue plasminogen activator (t-PA), and endothelial nitric oxide synthetase (eNOS).
- Compared proliferation and apoptosis rates across passages on both matrices.
Main Results:
- Early passages showed overexpression of prothrombotic factors (vWF, PAI) and underexpression of antithrombotic factors (t-PA, eNOS) on the model matrix.
- With increased passages on the model matrix, prothrombotic factor expression decreased while antithrombotic factor expression increased.
- Conventional matrices led to further amplification of prothrombotic factors and reduction of antithrombotic factors over passages; the model matrix supported better HSVEC proliferation and survival.
Conclusions:
- Matrix composition significantly influences the switching of EC phenotypes between prothrombotic and antithrombotic states.
- The biomimetic matrix model is suitable for studying exogenous factor effects on EC dysfunction in CVD contexts.
- This model offers a more physiologically relevant platform for investigating EC behavior in cardiovascular research.
Abstract:
The highly organized histological architecture of the vascular wall and the specialized cellular phenotypes are perturbed in conditions such as atherosclerosis, restenosis, and hypertension. Alterations of endothelial cell (EC) phenotype in cardiovascular diseases (CVDs) as an effect of alteration of extracellular matrix (ECM) composition have not been well understood. In vitro study of EC phenotype is limited because they tend to dedifferentiate in subcultures. The objective of this study was to use in vitro cell culture on a biomimetic matrix model to characterize phenotype of human saphenous vein endothelial cells (HSVECs) harvested from CVD patients. Parameters studied were mRNA expression and synthesis of von Willebrand factor (vWF), plasminogen activation inhibitor (PAI), tissue plasminogen activator (t-PA), and endothelial nitric oxide synthetase (eNOS). Proliferation and apoptosis of HSVEC cultures obtained from eight different patients were compared on two matrices until passage 12. In early passages, both the prothombotic molecules vWF and PAI were overexpressed, whereas the antithrombotic molecules t-PA and eNOS were underexpressed. With increase in passage number, low expression of prothrombotic molecules and high expression of antithrombotic molecules were seen in cells on the model matrix. But when cells were grown on conventional gelatin-coated polystyrene, expression of prothrombotic molecules amplified further and antithrombotic molecules lessened with the progression of passages. On the model matrix HSVECs showed good proliferation rate and survival through several passages. It is demonstrated that matrix composition can influence switching of EC phenotypes into pro/antithrombotic states. This matrix model may be suitable to study the effect of exogenous factors on EC dysfunction with respect to CVD.
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