Establishment of primary cultures of human brain microvascular endothelial cells: a new and simplified method to obtain cells for an in vitro model of the blood-brain barrier
| Autor: | Bernas, Michael J, Cardoso, Filipa L, Daley, Sarah K, Weinand, Martin E, Campos, Alexandre R, Ferreira, António J Gonçalves, Hoying, James B, Witte, Marlys H, Brites, Dora, Persidsky, Yuri, Ramirez, Servio H, Brito, Maria A |
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| Jazyk: | angličtina |
| Rok vydání: | 2010 |
| Předmět: |
Glucose Transporter Type 1
Caveolin 1 Cell Culture Techniques Endothelial Cells Hippocampus Models Biological Article Collagen Type I Temporal Lobe Tight Junctions Blood-Brain Barrier von Willebrand Factor Electric Impedance Humans ATP Binding Cassette Transporter Subfamily B Member 1 Pericytes Cells Cultured |
| Popis: | We describe a method for generating primary cultures of human brain microvascular endothelial cells (HBMVECs). HBMVECs are derived from microvessels isolated from temporal tissue removed during operative treatment of epilepsy. The tissue is mechanically fragmented and size filtered using polyester meshes. The resulting microvessel fragments are placed onto type I collagen-coated flasks to allow HBMVECs to migrate and proliferate. The overall process takes less than 3 h and does not require specialized equipment or enzymatic processes. HBMVECs are typically cultured for approximately 1 month until confluent. Cultures are highly pure ( approximately 97% endothelial cells; approximately 3% pericytes), are reproducible, and show characteristic brain endothelial markers (von Willebrand factor, glucose transporter-1) and robust expression of tight and adherens junction proteins as well as caveolin-1 and efflux protein P-glycoprotein. Monolayers of HBMVECs show characteristically high transendothelial electric resistance and have proven useful in multiple functional studies for in vitro modeling of the human blood-brain barrier. |
| Databáze: | OpenAIRE |
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