Abstract
Blood vessels are comprised of endothelial and smooth muscle cells. Obtaining both types of cells from vessels of living donors is not possible without invasive surgery. To address this, we have devised a strategy whereby human endothelial and smooth muscle cells derived from blood progenitors from the same donor could be cultured with autologous leukocytes to generate a same donor “vessel in a dish” bioassay. Autologous sets of blood outgrowth endothelial cells (BOECs), smooth muscle cells (BO‐SMCs), and leukocytes were obtained from four donors. Cells were treated in monoculture and cumulative coculture conditions. The endothelial specific mediator endothelin‐1 along with interleukin (IL)‐6, IL‐8, tumor necrosis factor α, and interferon gamma‐induced protein 10 were measured under control culture conditions and after stimulation with cytokines. Cocultures remained viable throughout. The profile of individual mediators released from cells was consistent with what we know of endothelial and smooth muscle cells cultured from blood vessels. For the first time, we report a proof of concept study where autologous blood outgrowth “vascular” cells and leukocytes were studied alone and in coculture. This novel bioassay has usefulness in vascular biology research, patient phenotyping, drug testing, and tissue engineering.
Highlights
Blood vessels are comprised of endothelial cells which line the luminal surface and the underlying smooth muscle cells which provide the structural component
We suggest that the use of blood outgrowth smooth muscle cells (BO-SMCs) along with autologous blood outgrowth endothelial cells (BOECs) and leukocytes can provide a comprehensive vascular bioassay system
Vascular smooth muscle cells release negligible levels of ET-1, unless stimulated with the specific combinations of IFN-γ plus tumor necrosis factor α (TNF-α).23,24. This feature of vascular smooth muscle cells distinguishes them from lung fibroblasts, which do not release ET-1 when stimulated with TNF-α and IFN-γ
Summary
Blood vessels are comprised of endothelial cells which line the luminal surface and the underlying smooth muscle cells which provide the structural component. From another, which introduces obvious confounding factors related to tissue mismatching and where HUVECs are used, immune privilege To address this for endothelial cells, we have developed a bioassay using BOECs in coculture with autologous leukocytes to generate a “same donor” platform for drug testing.[19] for a fuller understanding of drug action, patient phenotyping, and vascular inflammation, it is essential that the vascular smooth muscle component be considered. We suggest that the use of BO-SMCs along with autologous BOECs and leukocytes can provide a comprehensive vascular bioassay system This is important since without such an approach it is not possible to identify which cell type(s) (endothelial smooth muscle and/or leukocytes) are dysfunctional in vascular disease. Vasoactive and inflammatory mediators were measured under control and inflammatory conditions in culture
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