Novel flow cytometric analysis of the blood-brain barrier

Dionna W. Williams, Lydia Tesfa, Joan W. Berman

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

The blood-brain barrier (BBB) is primarily comprised of brain microvascular endothelial cells (BMVEC) and astrocytes and serves as a physical and chemical barrier that separates the periphery from the brain. We describe a flow cytometric method using our in vitro model of the human BBB to characterize BMVEC surface junctional proteins critical for maintenance of barrier function, cell viability, and leukocyte adhesion. For this methodology, BMVEC are cocultured with astrocytes in a transwell tissue culture insert to establish the barrier, after which time the BBB are treated with specific agents, and the BMVEC collected for flow cytometric analyses. We use a standard and optimized method to recover the BMVEC from the coculture model that maintains junctional protein expression and cell viability. A novel leukocyte adhesion assay enables a quantitative analysis of peripheral blood mononuclear cell (PBMC) interactions with the BMVEC and can be used to assess the adhesion of many cell types to the BBB. Furthermore, this method enables the concomitant analysis of a large number of adhesion molecules and tight junction proteins on both the BMVEC and adherent PBMC under homeostatic and pathologic conditions. Flow cytometry is an extremely powerful tool, and this technique can also be applied to assess variables not performed in this study, including cell cycle progression, and calcium flux.

Original languageEnglish (US)
Pages (from-to)897-907
Number of pages11
JournalCytometry Part A
Volume87
Issue number10
DOIs
StatePublished - Oct 1 2015

Fingerprint

Blood-Brain Barrier
Endothelial Cells
Brain
Astrocytes
Blood Cells
Cell Survival
Leukocytes
Tight Junction Proteins
Architectural Accessibility
Coculture Techniques
Cell Adhesion
Cell Communication
Cell Cycle
Flow Cytometry
Membrane Proteins
Maintenance
Calcium
Proteins

Keywords

  • Adhesion
  • Adhesion molecule
  • Brain microvascular endothelium
  • Flow cytometry
  • Leukocyte-endothelial cell interaction
  • Tight junction protein

ASJC Scopus subject areas

  • Cell Biology
  • Histology
  • Pathology and Forensic Medicine

Cite this

Novel flow cytometric analysis of the blood-brain barrier. / Williams, Dionna W.; Tesfa, Lydia; Berman, Joan W.

In: Cytometry Part A, Vol. 87, No. 10, 01.10.2015, p. 897-907.

Research output: Contribution to journalArticle

@article{7414ecff765442a0a6e80317a3e3adf7,
title = "Novel flow cytometric analysis of the blood-brain barrier",
abstract = "The blood-brain barrier (BBB) is primarily comprised of brain microvascular endothelial cells (BMVEC) and astrocytes and serves as a physical and chemical barrier that separates the periphery from the brain. We describe a flow cytometric method using our in vitro model of the human BBB to characterize BMVEC surface junctional proteins critical for maintenance of barrier function, cell viability, and leukocyte adhesion. For this methodology, BMVEC are cocultured with astrocytes in a transwell tissue culture insert to establish the barrier, after which time the BBB are treated with specific agents, and the BMVEC collected for flow cytometric analyses. We use a standard and optimized method to recover the BMVEC from the coculture model that maintains junctional protein expression and cell viability. A novel leukocyte adhesion assay enables a quantitative analysis of peripheral blood mononuclear cell (PBMC) interactions with the BMVEC and can be used to assess the adhesion of many cell types to the BBB. Furthermore, this method enables the concomitant analysis of a large number of adhesion molecules and tight junction proteins on both the BMVEC and adherent PBMC under homeostatic and pathologic conditions. Flow cytometry is an extremely powerful tool, and this technique can also be applied to assess variables not performed in this study, including cell cycle progression, and calcium flux.",
keywords = "Adhesion, Adhesion molecule, Brain microvascular endothelium, Flow cytometry, Leukocyte-endothelial cell interaction, Tight junction protein",
author = "Williams, {Dionna W.} and Lydia Tesfa and Berman, {Joan W.}",
year = "2015",
month = "10",
day = "1",
doi = "10.1002/cyto.a.22683",
language = "English (US)",
volume = "87",
pages = "897--907",
journal = "Cytometry. Part A : the journal of the International Society for Analytical Cytology",
issn = "1552-4922",
publisher = "Wiley-Liss Inc.",
number = "10",

}

TY - JOUR

T1 - Novel flow cytometric analysis of the blood-brain barrier

AU - Williams, Dionna W.

AU - Tesfa, Lydia

AU - Berman, Joan W.

PY - 2015/10/1

Y1 - 2015/10/1

N2 - The blood-brain barrier (BBB) is primarily comprised of brain microvascular endothelial cells (BMVEC) and astrocytes and serves as a physical and chemical barrier that separates the periphery from the brain. We describe a flow cytometric method using our in vitro model of the human BBB to characterize BMVEC surface junctional proteins critical for maintenance of barrier function, cell viability, and leukocyte adhesion. For this methodology, BMVEC are cocultured with astrocytes in a transwell tissue culture insert to establish the barrier, after which time the BBB are treated with specific agents, and the BMVEC collected for flow cytometric analyses. We use a standard and optimized method to recover the BMVEC from the coculture model that maintains junctional protein expression and cell viability. A novel leukocyte adhesion assay enables a quantitative analysis of peripheral blood mononuclear cell (PBMC) interactions with the BMVEC and can be used to assess the adhesion of many cell types to the BBB. Furthermore, this method enables the concomitant analysis of a large number of adhesion molecules and tight junction proteins on both the BMVEC and adherent PBMC under homeostatic and pathologic conditions. Flow cytometry is an extremely powerful tool, and this technique can also be applied to assess variables not performed in this study, including cell cycle progression, and calcium flux.

AB - The blood-brain barrier (BBB) is primarily comprised of brain microvascular endothelial cells (BMVEC) and astrocytes and serves as a physical and chemical barrier that separates the periphery from the brain. We describe a flow cytometric method using our in vitro model of the human BBB to characterize BMVEC surface junctional proteins critical for maintenance of barrier function, cell viability, and leukocyte adhesion. For this methodology, BMVEC are cocultured with astrocytes in a transwell tissue culture insert to establish the barrier, after which time the BBB are treated with specific agents, and the BMVEC collected for flow cytometric analyses. We use a standard and optimized method to recover the BMVEC from the coculture model that maintains junctional protein expression and cell viability. A novel leukocyte adhesion assay enables a quantitative analysis of peripheral blood mononuclear cell (PBMC) interactions with the BMVEC and can be used to assess the adhesion of many cell types to the BBB. Furthermore, this method enables the concomitant analysis of a large number of adhesion molecules and tight junction proteins on both the BMVEC and adherent PBMC under homeostatic and pathologic conditions. Flow cytometry is an extremely powerful tool, and this technique can also be applied to assess variables not performed in this study, including cell cycle progression, and calcium flux.

KW - Adhesion

KW - Adhesion molecule

KW - Brain microvascular endothelium

KW - Flow cytometry

KW - Leukocyte-endothelial cell interaction

KW - Tight junction protein

UR - http://www.scopus.com/inward/record.url?scp=84942374586&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84942374586&partnerID=8YFLogxK

U2 - 10.1002/cyto.a.22683

DO - 10.1002/cyto.a.22683

M3 - Article

VL - 87

SP - 897

EP - 907

JO - Cytometry. Part A : the journal of the International Society for Analytical Cytology

JF - Cytometry. Part A : the journal of the International Society for Analytical Cytology

SN - 1552-4922

IS - 10

ER -