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Updated: Jul 31, 2025
Human Saphenous Vein Endothelial Cell Isolation and Exposure to Controlled Levels of Shear Stress and Stretch
Published on: April 21, 2023
Human Saphenous Vein Endothelial Cell Isolation and Exposure to Controlled Levels of Shear Stress and Stretch
Thaís Girão-Silva1, Miriam Helena Fonseca-Alaniz2, Luís Alberto Oliveira Dallan2
1Instituto do Coraçao (InCor), Hospital das Clinicas HCFMUSP, Faculdade de Medicina, Universidade de Sao Paulo; Cardiology Research Group, Faculty VI Medicine and Health Sciences, Carl von Ossietzky University of Oldenburg.
Insights
Researchers isolated human saphenous vein endothelial cells (hSVECs) to study how hemodynamic stress affects graft patency. Mechanical forces like shear stress and stretch alter hSVEC behavior, providing insights into saphenous vein graft performance.
Area of Science:
- Cardiovascular Biology
- Vascular Cell Biology
- Biomedical Engineering
Background:
- Coronary artery bypass graft (CABG) surgery uses saphenous vein grafts (SVGs), but their long-term patency is limited compared to arterial grafts.
- Hemodynamic stress during arterialization of SVGs can cause endothelial damage, potentially explaining reduced graft patency.
Purpose of the Study:
- To isolate, characterize, and expand human saphenous vein endothelial cells (hSVECs).
- To investigate the effects of mechanical stress (shear and stretch) on hSVECs, mimicking conditions within arterialized SVGs.
Main Methods:
- hSVECs were isolated using collagenase digestion and characterized by morphology and endothelial markers (CD31, VE-cadherin).
- Cells were subjected to shear stress in a parallel plate flow chamber.
- Cells were subjected to mechanical stretch using a silicon membrane system to simulate venous and arterial conditions.
Main Results:
- Isolated hSVECs exhibited typical endothelial morphology and marker expression.
- Shear stress induced cell alignment and altered expression of KLF2, KLF4, and NOS3.
- Mechanical stretch modulated F-actin organization and nitric oxide (NO) secretion in hSVECs.
Conclusions:
- A detailed method for isolating and culturing hSVECs was established.
- Hemodynamic mechanical stresses significantly influence hSVEC phenotype and function.
- This model provides a platform for studying endothelial responses to mechanical forces in SVGs.
Abstract:
Coronary artery bypass graft (CABG) surgery is a procedure to revascularize ischemic myocardium. Saphenous vein remains used as a CABG conduit despite the reduced long-term patency compared to arterial conduits. The abrupt increase of hemodynamic stress associated with the graft arterialization results in vascular damage, especially the endothelium, that may influence the low patency of the saphenous vein graft (SVG). Here, we describe the isolation, characterization, and expansion of human saphenous vein endothelial cells (hSVECs). Cells isolated by collagenase digestion display the typical cobblestone morphology and express endothelial cell markers CD31 and VE-cadherin. To assess the mechanical stress influence, protocols were used in this study to investigate the two main physical stimuli, shear stress and stretch, on arterialized SVGs. hSVECs are cultured in a parallel plate flow chamber to produce shear stress, showing alignment in the direction of the flow and increased expression of KLF2, KLF4, and NOS3. hSVECs can also be cultured in a silicon membrane that allows controlled cellular stretch mimicking venous (low) and arterial (high) stretch. Endothelial cells' F-actin pattern and nitric oxide (NO) secretion are modulated accordingly by the arterial stretch. In summary, we present a detailed method to isolate hSVECs to study the influence of hemodynamic mechanical stress on an endothelial phenotype.
