Immortalized human cerebral microvascular endothelial cells maintain the properties of primary cells in an in vitro model of immune migration across the blood brain barrier

Brian P. Daniels, Lillian Cruz-Orengo, Tracy Jo Pasieka, Pierre Olivier Couraud, Ignacio A. Romero, Babette Weksler, John A. Cooper, Tamara L. Doering, Robyn S. Klein

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

The immortalized human cerebral microvascular endothelial cell line HCMEC/D3 presents a less expensive and more logistically feasible alternative to primary human brain microvascular endothelial cells (HBMEC's) for use in constructing in vitro models of the blood brain barrier (BBB). However, the fidelity of the HCMEC/D3 cell line to primary HBMEC's in studies of immune transmigration has yet to be established. Flow cytometric analysis of primary human leukocyte migration across in vitro BBB's generated with either HCMEC/D3 or primary HBMEC's revealed that HCMEC/D3 maintains the immune barrier properties of primary HBMEC's. Leukocyte migration responses and inflammatory cytokine production were statistically indistinguishable between both endothelial cell types, and both cell types responded similarly to astrocyte coculture, stimulation of leukocytes with phorbol myristate acetate (PMA) and ionomycin, and inflammatory cytokine treatment. This report is the first to validate the HCMEC/D3 cell line in a neuroimmunological experimental system via direct comparison to primary HBMEC's, demonstrating remarkable fidelity in terms of barrier resistance, immune migration profiles, and responsiveness to inflammatory cytokines. Moreover, we report novel findings demonstrating that interaction effects between immune cells and resident CNS cells are preserved in HCMEC/D3, suggesting that important characteristics of neuroimmune interactions during CNS inflammation are preserved in systems utilizing this cell line. Together, these findings demonstrate that HCMEC/D3 is a valid and powerful tool for less expensive and higher throughput in vitro investigations of immune migration at the BBB.

Original languageEnglish (US)
Pages (from-to)173-179
Number of pages7
JournalJournal of Neuroscience Methods
Volume212
Issue number1
DOIs
StatePublished - Jan 1 2013
Externally publishedYes

Fingerprint

Blood-Brain Barrier
Endothelial Cells
Brain
Cell Line
Leukocytes
Cytokines
Ionomycin
Tetradecanoylphorbol Acetate
Coculture Techniques
In Vitro Techniques
Astrocytes
Inflammation

Keywords

  • Blood brain barrier
  • HCMEC/D3
  • Immune trafficking
  • In vitro model

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Immortalized human cerebral microvascular endothelial cells maintain the properties of primary cells in an in vitro model of immune migration across the blood brain barrier. / Daniels, Brian P.; Cruz-Orengo, Lillian; Pasieka, Tracy Jo; Couraud, Pierre Olivier; Romero, Ignacio A.; Weksler, Babette; Cooper, John A.; Doering, Tamara L.; Klein, Robyn S.

In: Journal of Neuroscience Methods, Vol. 212, No. 1, 01.01.2013, p. 173-179.

Research output: Contribution to journalArticle

Daniels, Brian P. ; Cruz-Orengo, Lillian ; Pasieka, Tracy Jo ; Couraud, Pierre Olivier ; Romero, Ignacio A. ; Weksler, Babette ; Cooper, John A. ; Doering, Tamara L. ; Klein, Robyn S. / Immortalized human cerebral microvascular endothelial cells maintain the properties of primary cells in an in vitro model of immune migration across the blood brain barrier. In: Journal of Neuroscience Methods. 2013 ; Vol. 212, No. 1. pp. 173-179.
@article{36088bd7c33f40238d75149d6027096e,
title = "Immortalized human cerebral microvascular endothelial cells maintain the properties of primary cells in an in vitro model of immune migration across the blood brain barrier",
abstract = "The immortalized human cerebral microvascular endothelial cell line HCMEC/D3 presents a less expensive and more logistically feasible alternative to primary human brain microvascular endothelial cells (HBMEC's) for use in constructing in vitro models of the blood brain barrier (BBB). However, the fidelity of the HCMEC/D3 cell line to primary HBMEC's in studies of immune transmigration has yet to be established. Flow cytometric analysis of primary human leukocyte migration across in vitro BBB's generated with either HCMEC/D3 or primary HBMEC's revealed that HCMEC/D3 maintains the immune barrier properties of primary HBMEC's. Leukocyte migration responses and inflammatory cytokine production were statistically indistinguishable between both endothelial cell types, and both cell types responded similarly to astrocyte coculture, stimulation of leukocytes with phorbol myristate acetate (PMA) and ionomycin, and inflammatory cytokine treatment. This report is the first to validate the HCMEC/D3 cell line in a neuroimmunological experimental system via direct comparison to primary HBMEC's, demonstrating remarkable fidelity in terms of barrier resistance, immune migration profiles, and responsiveness to inflammatory cytokines. Moreover, we report novel findings demonstrating that interaction effects between immune cells and resident CNS cells are preserved in HCMEC/D3, suggesting that important characteristics of neuroimmune interactions during CNS inflammation are preserved in systems utilizing this cell line. Together, these findings demonstrate that HCMEC/D3 is a valid and powerful tool for less expensive and higher throughput in vitro investigations of immune migration at the BBB.",
keywords = "Blood brain barrier, HCMEC/D3, Immune trafficking, In vitro model",
author = "Daniels, {Brian P.} and Lillian Cruz-Orengo and Pasieka, {Tracy Jo} and Couraud, {Pierre Olivier} and Romero, {Ignacio A.} and Babette Weksler and Cooper, {John A.} and Doering, {Tamara L.} and Klein, {Robyn S.}",
year = "2013",
month = "1",
day = "1",
doi = "10.1016/j.jneumeth.2012.10.001",
language = "English (US)",
volume = "212",
pages = "173--179",
journal = "Journal of Neuroscience Methods",
issn = "0165-0270",
publisher = "Elsevier",
number = "1",

}

TY - JOUR

T1 - Immortalized human cerebral microvascular endothelial cells maintain the properties of primary cells in an in vitro model of immune migration across the blood brain barrier

AU - Daniels, Brian P.

AU - Cruz-Orengo, Lillian

AU - Pasieka, Tracy Jo

AU - Couraud, Pierre Olivier

AU - Romero, Ignacio A.

AU - Weksler, Babette

AU - Cooper, John A.

AU - Doering, Tamara L.

AU - Klein, Robyn S.

PY - 2013/1/1

Y1 - 2013/1/1

N2 - The immortalized human cerebral microvascular endothelial cell line HCMEC/D3 presents a less expensive and more logistically feasible alternative to primary human brain microvascular endothelial cells (HBMEC's) for use in constructing in vitro models of the blood brain barrier (BBB). However, the fidelity of the HCMEC/D3 cell line to primary HBMEC's in studies of immune transmigration has yet to be established. Flow cytometric analysis of primary human leukocyte migration across in vitro BBB's generated with either HCMEC/D3 or primary HBMEC's revealed that HCMEC/D3 maintains the immune barrier properties of primary HBMEC's. Leukocyte migration responses and inflammatory cytokine production were statistically indistinguishable between both endothelial cell types, and both cell types responded similarly to astrocyte coculture, stimulation of leukocytes with phorbol myristate acetate (PMA) and ionomycin, and inflammatory cytokine treatment. This report is the first to validate the HCMEC/D3 cell line in a neuroimmunological experimental system via direct comparison to primary HBMEC's, demonstrating remarkable fidelity in terms of barrier resistance, immune migration profiles, and responsiveness to inflammatory cytokines. Moreover, we report novel findings demonstrating that interaction effects between immune cells and resident CNS cells are preserved in HCMEC/D3, suggesting that important characteristics of neuroimmune interactions during CNS inflammation are preserved in systems utilizing this cell line. Together, these findings demonstrate that HCMEC/D3 is a valid and powerful tool for less expensive and higher throughput in vitro investigations of immune migration at the BBB.

AB - The immortalized human cerebral microvascular endothelial cell line HCMEC/D3 presents a less expensive and more logistically feasible alternative to primary human brain microvascular endothelial cells (HBMEC's) for use in constructing in vitro models of the blood brain barrier (BBB). However, the fidelity of the HCMEC/D3 cell line to primary HBMEC's in studies of immune transmigration has yet to be established. Flow cytometric analysis of primary human leukocyte migration across in vitro BBB's generated with either HCMEC/D3 or primary HBMEC's revealed that HCMEC/D3 maintains the immune barrier properties of primary HBMEC's. Leukocyte migration responses and inflammatory cytokine production were statistically indistinguishable between both endothelial cell types, and both cell types responded similarly to astrocyte coculture, stimulation of leukocytes with phorbol myristate acetate (PMA) and ionomycin, and inflammatory cytokine treatment. This report is the first to validate the HCMEC/D3 cell line in a neuroimmunological experimental system via direct comparison to primary HBMEC's, demonstrating remarkable fidelity in terms of barrier resistance, immune migration profiles, and responsiveness to inflammatory cytokines. Moreover, we report novel findings demonstrating that interaction effects between immune cells and resident CNS cells are preserved in HCMEC/D3, suggesting that important characteristics of neuroimmune interactions during CNS inflammation are preserved in systems utilizing this cell line. Together, these findings demonstrate that HCMEC/D3 is a valid and powerful tool for less expensive and higher throughput in vitro investigations of immune migration at the BBB.

KW - Blood brain barrier

KW - HCMEC/D3

KW - Immune trafficking

KW - In vitro model

UR - http://www.scopus.com/inward/record.url?scp=84868249383&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84868249383&partnerID=8YFLogxK

U2 - 10.1016/j.jneumeth.2012.10.001

DO - 10.1016/j.jneumeth.2012.10.001

M3 - Article

C2 - 23068604

AN - SCOPUS:84868249383

VL - 212

SP - 173

EP - 179

JO - Journal of Neuroscience Methods

JF - Journal of Neuroscience Methods

SN - 0165-0270

IS - 1

ER -