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Updated: Oct 27, 2025
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Microvascular and Macrovascular Endothelial Cell Isolation and Purification from Lung-Derived Samples
Published on: February 3, 2023
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Isolation of Microvascular Endothelial Cells.
Kenneth C P Cheung1, Federica M Marelli-Berg1
1William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London EC1M 6BQ, United Kingdom.
Bio-Protocol
|July 21, 2021
Summary
Researchers developed a fast, efficient method to isolate murine microvascular endothelial cells for cardiovascular research. This technique uses standard lab equipment and magnetic bead separation for high-purity cell cultures.
Area of Science:
- Cardiovascular Biology
- Cell Biology
- Biotechnology
Background:
- Vascular endothelial cells maintain vascular homeostasis.
- Endothelial dysfunction contributes to cardiovascular diseases.
- Murine endothelial cell culture is vital for cardiovascular research.
Purpose of the Study:
- To present a rapid and efficient protocol for isolating microvascular endothelial cells from murine tissues.
- To enable cardiovascular disease research using readily available laboratory equipment.
Main Methods:
- Mechanical mincing and enzymatic digestion (collagenase, trypsin) of murine lung or heart tissue.
- Incubation of single-cell suspension with CD31, CD105 antibodies, and biotinylated isolectin B-4.
- Magnetic bead separation using anti-mouse Ig- and streptavidin-conjugated microbeads for endothelial cell isolation.
Main Results:
- Successful isolation and expansion of murine microvascular endothelial cells.
- Stable morphological and phenotypic characteristics maintained over 10 passages.
- Absence of contaminating non-endothelial cells throughout the process.
Conclusions:
- This protocol provides a quick, efficient, and equipment-independent method for isolating high-purity murine microvascular endothelial cells.
- The isolated cells are suitable for various downstream analyses in cardiovascular research.
- The method ensures the integrity and purity of endothelial cell cultures for reliable experimental outcomes.
