Home Neuroscience Isolation of Primary Murine Brain Microvascular Endothelial Cells
Neuroscience JoVE (Open Access) Citable · DOI

Isolation of Primary Murine Brain Microvascular Endothelial Cells

DOI: 10.3791/52204-v
What you'll learn
  • Isolate primary murine brain microvascular endothelial cells from mouse brain tissue
  • Characterize morphological and functional properties of cultured BMECs in vitro
  • Establish in vitro blood-brain-barrier model for experimental studies
Protocol

Brain microvascular endothelial cells (BMEC) are interconnected by specific junctional proteins forming a highly regulated barrier separating blood and the central nervous system (CNS), the so-called blood-brain-barrier (BBB). The isolation of primary murine brain microvascular endothelial cells, as discussed in this protocol, enables detailed in vitro studies of the BBB.

Difficulty
advanced
Total time
~4–6 hours per mouse (tissue isolation through initial culture)
Model organism
Mouse (strain unspecified)
Biosafety
BSL-1

Steps

1
Isolate murine brain microvascular endothelial cells

Extract primary BMECs from mouse brain tissue using enzymatic digestion and mechanical separation. The protocol yields pure, viable endothelial cells suitable for in vitro BBB studies.

▶ 01:46
2
Assess morphological and functional BMEC properties

Verify isolated cells display characteristic endothelial morphology and barrier function properties, including tight junction protein expression and transepithelial electrical resistance (TEER) measurements.

▶ 05:41
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