Isolation and expansion of human and mouse brain microvascular endothelial cells

Stefania E. Navone, Giovanni Marfia, Gloria Invernici, Silvia Cristini, Sara Nava, Sergio Balbi, Simone Sangiorgi, Emilio Ciusani, Alessandra Bosutti, Giulio Alessandri, Mark Slevin, Eugenio A. Parati

Research output: Contribution to journalArticlepeer-review


Brain microvascular endothelial cells (BMVECs) have an important role in the constitution of the blood-brain barrier (BBB). The BBB is involved in the disease processes of a number of neurological disorders in which its permeability increases. Isolation of BMVECs could elucidate the mechanism involved in these processes. This protocol describes how to isolate and expand human and mouse BMVECs. The procedure covers brain-tissue dissociation, digestion and cell selection. Cells are selected on the basis of time-responsive differential adhesiveness to a collagen type I-precoated surface. The protocol also describes immunophenotypic characterization, cord formation and functional assays to confirm that these cells in endothelial proliferation medium (EndoPM) have an endothelial origin. The entire technique requires ∼7 h of active time. Endothelial cell clusters are readily visible after 48 h, and expansion of BMVECs occurs over the course of ∼60 d.

Original languageEnglish
Pages (from-to)1680-1693
Number of pages14
JournalNature Protocols
Issue number9
Publication statusPublished - Sep 2013

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)


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