Primary Porcine Brain Microvessel Endothelial Cell Isolation and Culture. Issue 1 (8th October 2014)

• Record Type: Journal Article
• Authors: Patabendige, Adjanie; Abbott, N. Joan
• Editors: Gerfen, Charles R.; Holmes, Andrew; Sibley, David; Skolnick, Phil; Wray, Susan
• Is Part Of: Current protocols in neuroscience. Issue 69:Issue 1(2014)
• Page Start: 3.27.1
• Page End: 3.27.17
• Publication Date: 2014-10-08
• Subjects: blood‐brain barrier -- porcine brain endothelial cells -- transendothelial electrical resistance -- cell culture -- astrocyte‐conditioned medium; Neurosciences -- Laboratory manuals; Neurosciences; Cytological Techniques; Histological Techniques; Nervous System -- anatomy & histology; Neurosciences; Laboratory Manual; Laboratory manuals; 612.8
• DOI: 10.1002/0471142301.ns0327s69
• Languages: English
• ISSNs: 1934-8576

Abstract

ABSTRACT: Cell culture models of the blood‐brain barrier (BBB) are useful tools to study the functionality of the BBB in health and disease. Several good in vitro BBB models are available from different species. However, most brain endothelial cells lose some of their in vivo BBB phenotype in culture. Porcine brain endothelial cells (PBECs) tend to retain most of their in vivo BBB characteristics and usually give higher transendothelial electrical resistance (TEER, representing functional well‐developed tight junctions) compared to brain endothelial cells from other species. The protocol described in this unit gives detailed instructions for isolation and culture of PBECs from fresh porcine brains. This porcine BBB model generates high TEER without the need for co‐culture with astrocytes. However, astrocyte‐derived factors can be introduced to the system through the use of astrocyte‐conditioned medium or co‐culture with astrocytes, which may be necessary for further enhancing the BBB phenotype for certain complex studies. Curr. Protoc. Neurosci . 69:3.27.1‐3.27.17. © 2014 by John Wiley & Sons, Inc.