Methods to Assess Pericyte-Endothelial Cell Interactions in a Coculture Model

Abstract

The blood-brain barrier (BBB) comprises the microvascular endothelial cells, pericytes, and astrocytes, which are connected by the extracellular matrix (ECM). Current BBB models focus solely on the microvascular endothelial cells which constitute a physical barrier by formation of tight junctions (TJs), while the impact of pericytes on barrier regulation is poorly understood. We established a coculture model from primary porcine brain capillary endothelial cells (PBCECs) and pericytes (PBCPs) to approach the in vivo situation. This model allows the examination of pericyte impact on pharmacological, transport, migration, and metabolic activity of the BBB. In vivo the interaction between pericytes and endothelial cells is partly controlled by the ECM which is remodeled by matrix metalloproteinases (MMPs). Both endothelial cells and pericytes secrete MMPs which are important not only for ECM remodeling but also for TJ cleavage. In this chapter, current methods to study the interactions of these cell types by ECM signaling as well as MMP secretion are described.

Development, maintenance and disruption of the blood-brain barrier

“…A recent innovative culture system allows human pluripotent stem cells to co-differentiate into cerebrovascular ECs and neural progenitors that signal to ECs and thereby promote the establishment of relevant BBB properties 148 . Furthermore, co-culture systems are becoming increasingly popular as they account for the importance of pericytes and astrocytes in the regulation of the BBB 149,150 . Even more complex triple culture systems have been explored, and microglia as well as neurons have been included in such systems 151-154 .…”

Vascular Calcification: Is it rather a Stem/Progenitor Cells Driven Phenomenon?

“…They facilitate and integrate cell communication (Armulik et al, 2005 ). Usually, pericytes overlap several ECs and regulate certain functions by secretion of factors (Armulik et al, 2005 ; Thanabalasundaram et al, 2011 ). Pericytes extend long processes exhibiting contractile microfilament bundles which wrap around the blood vessel (Dore-Duffy and Cleary, 2011 ).…”

Blood-Brain Barrier Effects of the Fusarium Mycotoxins Deoxynivalenol, 3 Acetyldeoxynivalenol, and Moniliformin and Their Transfer to the Brain

“…Primary porcine brain capillary endothelial cells (PBCEC) form a well characterized, flexible model to study interactions of pericytes [ 6 ] and astrocytes [ 7 ] with endothelial cells. Furthermore, they have been successfully used for transport studies of xenobiotics [ 8 , 9 ].…”