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Updated: Aug 5, 2026
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Isolation of Primary Murine Brain Microvascular Endothelial Cells
Published on: November 14, 2014
Isolation and Culture of Mouse Brain Microvascular Endothelial Cells
Esthefanny Escudero-Guevara1, Hermes Sandoval1, Jesenia Acurio2
1Department of Basic Sciences, Vascular Physiology Laboratory; Biomedical Sciences Doctorate Program, Universidad de Talca.
Journal of Visualized Experiments : Jove
|August 3, 2026
Summary
This study presents a reliable protocol for isolating and culturing primary brain microvascular endothelial cells (BMECs) from mouse brains. These BMEC cultures are crucial for studying the blood-brain barrier (BBB) and neurological diseases.
Area of Science:
- Neuroscience
- Cell Biology
- Biomedical Engineering
Background:
- Brain microvascular endothelial cells (BMECs) form the blood-brain barrier (BBB), regulating CNS homeostasis.
- In vitro models of brain endothelium are vital for studying BBB function and disease.
- Existing methods may lack reproducibility for obtaining primary BMEC cultures.
Purpose of the Study:
- To describe a reproducible protocol for isolating and culturing primary BMECs from neonatal murine cerebral cortex.
- To provide a reliable method for generating BMEC cultures for research.
Main Methods:
- Neonatal murine cerebral cortex dissection and dissociation (mechanical and enzymatic).
- Bovine serum albumin (BSA) density gradient for myelin debris removal.
- CD31-based immunoselection for endothelial cell enrichment.
- Culture on collagen-coated plates and characterization.
Main Results:
- Successful isolation and primary culture of BMECs.
- Characterization via morphology, immunofluorescence (endothelial markers), tube formation assays.
- Evaluation of barrier properties using transendothelial electrical resistance (TEER).
Conclusions:
- The described protocol offers a reliable approach for obtaining primary BMEC cultures.
- These cultures are suitable for investigating BBB biology, endothelial signaling, and neurovascular interactions.
- This method supports research in physiological and pathological conditions affecting the CNS.
