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Published on: September 9, 2016
Cultivation and characterization of micro- and macrovascular endothelial cells from the human heart
M Gräfe1, K Graf, W Auch-Schwelk
1Department of Cardiology, German Heart Institute Berlin.
Insights
Researchers developed a new method to isolate pure human heart endothelial cells for studying diseases like atherosclerosis. This technique ensures high purity, enabling reliable in vitro research on cardiovascular conditions.
Area of Science:
- Cardiovascular Biology
- Cell Biology
- Biotechnology
Background:
- Investigating cardiovascular diseases like atherosclerosis and inflammation requires pure human endothelial cell cultures.
- Human heart endothelial cells are crucial for in vitro disease modeling.
Purpose of the Study:
- To establish a reliable method for isolating pure viable endothelial cells from human hearts.
- To enable in vitro studies of cardiovascular processes.
Main Methods:
- Endothelial cells were isolated from human heart micro- and macrovasculature.
- A novel technique using Ulex Europaeus I (UEA-I) lectin-conjugated paramagnetic beads was employed for cell selection.
- Purity was assessed via immunological staining and fluorescence-activated cell sorting (FACS).
Main Results:
- The developed method yielded highly pure endothelial cell cultures with less than 2% contamination.
- Isolated cells exhibited typical endothelial markers (Factor VIII-related antigen, CD31, E-selectin).
- Cells retained key endothelial functions, including Dil-Ac-LDL uptake, angiotensin-converting enzyme activity, and tissue plasminogen activator secretion.
Conclusions:
- A robust method for obtaining pure human heart endothelial cells was established.
- These cells maintain essential endothelial characteristics and functions in culture.
- The method supports advanced in vitro research into atherosclerosis and inflammation.
Abstract:
In order to investigate processes, such as atherosclerosis and inflammation in vitro, it is necessary to obtain viable and pure endothelial cell cultures from human hearts. To this end, endothelial cells were isolated and cultured from the micro- and macrovasculature of human hearts obtained during heart transplantation. Isolation of capillaries after enzymatic digestion of heart muscle provided a source of microvascular endothelial cells. Contaminating non-endothelial cells were removed by a new technique: paramagnetic beads linked to the lectin ulex europaeus I (UEA-I) were used to select endothelial cells. The resulting cultures contained less than 2% of non-endothelial cells, as judged from immunological staining and fluorescence-activated cell sorting. Both types of endothelial cell displayed typical endothelial properties. They were all positive for factor VIII-related antigen and expressed the endothelial-specific adhesion molecules, CD31 and E-selectin (ELAM-1), after stimulation with cytokines. In addition, they could be labelled with Dil-Ac-LDL, contained angiotensin converting enzyme activity and secreted tissue plasminogen activator, thus demonstrating that typical endothelial functions were preserved in culture.
