Expression and deposition of basement membrane proteins by brain capillary endothelial cells in a primary murine model of the blood-brain barrier

Research output: Contribution to book/anthology/report/conference proceedingConference abstract in proceedingResearchpeer-review

Abstract

The blood-brain barrier (BBB) represents the interface between the blood and the brain parenchyma and consists of endothelial cells which are tightly sealed together by tight junction proteins. The endothelial cells are in addition supported by pericytes, which are embedded in the vascular basement membrane, and astrocyte endfeet. To study the interaction of the different cells of the BBB, construction of in vitro BBB models is valuable. However, the modulation and contribution of the cells of the BBB to the synthesis of basement membrane proteins in vitro is not fully elaborated. Thus, the aim of the present study was to create four different in vitro constructs of the murine BBB to characterise if the expression and secretion of basement membrane proteins by the murine brain capillary endothelial cells (mBCECs) was affected by co-culturing with pericytes, mixed glial cells, or both. Primary mBCECs and pericytes were isolated from brains of adult mice. Mixed glial cells were
prepared from cerebral cortices of newborn mice. The mBCECs were grown as mono-culture, or co-cultured with pericytes, mixed glial cells, or both. To study the expression of basement membrane proteins RT-qPCR, mass spectrometry, and immunofluorescent labelling were used. The mBCECs were found to express major basement membrane proteins in vitro and increased expression of laminin α5 and collagen IV α1 was correlated to the addition of BBB inducing factors (hydrocortisone, Ro20-1724, pCPT-cAMP). Co-culturing of the mBCECs with pericytes, mixed glial cells or both did not influence the gene expression of the investigated basement membrane proteins.
Original languageEnglish
Title of host publicationFinal Programme, 19th International Symposium on Signal Transduction at the Blood-Brain Barriers, 14-16 September 2016, Copenhagen, Denmark
PublisherUniversity of Copenhagen
Publication date2016
Pages37
Article numberP-12
Publication statusPublished - 2016
Event19th International Symposium on Signal Transduction at the Blood-Brain Barriers - København, Denmark
Duration: 14 Sep 201616 Sep 2016

Conference

Conference19th International Symposium on Signal Transduction at the Blood-Brain Barriers
CountryDenmark
CityKøbenhavn
Period14/09/201616/09/2016

Cite this

Thomsen, M. S., Birkelund, S., Larsen, A. B., Stensballe, A., & Moos, T. (2016). Expression and deposition of basement membrane proteins by brain capillary endothelial cells in a primary murine model of the blood-brain barrier. In Final Programme, 19th International Symposium on Signal Transduction at the Blood-Brain Barriers, 14-16 September 2016, Copenhagen, Denmark (pp. 37). [P-12] University of Copenhagen.
Thomsen, Maj Schneider ; Birkelund, Svend ; Larsen, Annette Burkhart ; Stensballe, Allan ; Moos, Torben. / Expression and deposition of basement membrane proteins by brain capillary endothelial cells in a primary murine model of the blood-brain barrier. Final Programme, 19th International Symposium on Signal Transduction at the Blood-Brain Barriers, 14-16 September 2016, Copenhagen, Denmark. University of Copenhagen, 2016. pp. 37
@inbook{88ad64910e9945b0aeaeb77356aee544,
title = "Expression and deposition of basement membrane proteins by brain capillary endothelial cells in a primary murine model of the blood-brain barrier",
abstract = "The blood-brain barrier (BBB) represents the interface between the blood and the brain parenchyma and consists of endothelial cells which are tightly sealed together by tight junction proteins. The endothelial cells are in addition supported by pericytes, which are embedded in the vascular basement membrane, and astrocyte endfeet. To study the interaction of the different cells of the BBB, construction of in vitro BBB models is valuable. However, the modulation and contribution of the cells of the BBB to the synthesis of basement membrane proteins in vitro is not fully elaborated. Thus, the aim of the present study was to create four different in vitro constructs of the murine BBB to characterise if the expression and secretion of basement membrane proteins by the murine brain capillary endothelial cells (mBCECs) was affected by co-culturing with pericytes, mixed glial cells, or both. Primary mBCECs and pericytes were isolated from brains of adult mice. Mixed glial cells wereprepared from cerebral cortices of newborn mice. The mBCECs were grown as mono-culture, or co-cultured with pericytes, mixed glial cells, or both. To study the expression of basement membrane proteins RT-qPCR, mass spectrometry, and immunofluorescent labelling were used. The mBCECs were found to express major basement membrane proteins in vitro and increased expression of laminin α5 and collagen IV α1 was correlated to the addition of BBB inducing factors (hydrocortisone, Ro20-1724, pCPT-cAMP). Co-culturing of the mBCECs with pericytes, mixed glial cells or both did not influence the gene expression of the investigated basement membrane proteins.",
author = "Thomsen, {Maj Schneider} and Svend Birkelund and Larsen, {Annette Burkhart} and Allan Stensballe and Torben Moos",
year = "2016",
language = "English",
pages = "37",
booktitle = "Final Programme, 19th International Symposium on Signal Transduction at the Blood-Brain Barriers, 14-16 September 2016, Copenhagen, Denmark",
publisher = "University of Copenhagen",

}

Thomsen, MS, Birkelund, S, Larsen, AB, Stensballe, A & Moos, T 2016, Expression and deposition of basement membrane proteins by brain capillary endothelial cells in a primary murine model of the blood-brain barrier. in Final Programme, 19th International Symposium on Signal Transduction at the Blood-Brain Barriers, 14-16 September 2016, Copenhagen, Denmark., P-12, University of Copenhagen, pp. 37, København, Denmark, 14/09/2016.

Expression and deposition of basement membrane proteins by brain capillary endothelial cells in a primary murine model of the blood-brain barrier. / Thomsen, Maj Schneider; Birkelund, Svend; Larsen, Annette Burkhart; Stensballe, Allan; Moos, Torben.

Final Programme, 19th International Symposium on Signal Transduction at the Blood-Brain Barriers, 14-16 September 2016, Copenhagen, Denmark. University of Copenhagen, 2016. p. 37 P-12.

Research output: Contribution to book/anthology/report/conference proceedingConference abstract in proceedingResearchpeer-review

TY - ABST

T1 - Expression and deposition of basement membrane proteins by brain capillary endothelial cells in a primary murine model of the blood-brain barrier

AU - Thomsen, Maj Schneider

AU - Birkelund, Svend

AU - Larsen, Annette Burkhart

AU - Stensballe, Allan

AU - Moos, Torben

PY - 2016

Y1 - 2016

N2 - The blood-brain barrier (BBB) represents the interface between the blood and the brain parenchyma and consists of endothelial cells which are tightly sealed together by tight junction proteins. The endothelial cells are in addition supported by pericytes, which are embedded in the vascular basement membrane, and astrocyte endfeet. To study the interaction of the different cells of the BBB, construction of in vitro BBB models is valuable. However, the modulation and contribution of the cells of the BBB to the synthesis of basement membrane proteins in vitro is not fully elaborated. Thus, the aim of the present study was to create four different in vitro constructs of the murine BBB to characterise if the expression and secretion of basement membrane proteins by the murine brain capillary endothelial cells (mBCECs) was affected by co-culturing with pericytes, mixed glial cells, or both. Primary mBCECs and pericytes were isolated from brains of adult mice. Mixed glial cells wereprepared from cerebral cortices of newborn mice. The mBCECs were grown as mono-culture, or co-cultured with pericytes, mixed glial cells, or both. To study the expression of basement membrane proteins RT-qPCR, mass spectrometry, and immunofluorescent labelling were used. The mBCECs were found to express major basement membrane proteins in vitro and increased expression of laminin α5 and collagen IV α1 was correlated to the addition of BBB inducing factors (hydrocortisone, Ro20-1724, pCPT-cAMP). Co-culturing of the mBCECs with pericytes, mixed glial cells or both did not influence the gene expression of the investigated basement membrane proteins.

AB - The blood-brain barrier (BBB) represents the interface between the blood and the brain parenchyma and consists of endothelial cells which are tightly sealed together by tight junction proteins. The endothelial cells are in addition supported by pericytes, which are embedded in the vascular basement membrane, and astrocyte endfeet. To study the interaction of the different cells of the BBB, construction of in vitro BBB models is valuable. However, the modulation and contribution of the cells of the BBB to the synthesis of basement membrane proteins in vitro is not fully elaborated. Thus, the aim of the present study was to create four different in vitro constructs of the murine BBB to characterise if the expression and secretion of basement membrane proteins by the murine brain capillary endothelial cells (mBCECs) was affected by co-culturing with pericytes, mixed glial cells, or both. Primary mBCECs and pericytes were isolated from brains of adult mice. Mixed glial cells wereprepared from cerebral cortices of newborn mice. The mBCECs were grown as mono-culture, or co-cultured with pericytes, mixed glial cells, or both. To study the expression of basement membrane proteins RT-qPCR, mass spectrometry, and immunofluorescent labelling were used. The mBCECs were found to express major basement membrane proteins in vitro and increased expression of laminin α5 and collagen IV α1 was correlated to the addition of BBB inducing factors (hydrocortisone, Ro20-1724, pCPT-cAMP). Co-culturing of the mBCECs with pericytes, mixed glial cells or both did not influence the gene expression of the investigated basement membrane proteins.

M3 - Conference abstract in proceeding

SP - 37

BT - Final Programme, 19th International Symposium on Signal Transduction at the Blood-Brain Barriers, 14-16 September 2016, Copenhagen, Denmark

PB - University of Copenhagen

ER -

Thomsen MS, Birkelund S, Larsen AB, Stensballe A, Moos T. Expression and deposition of basement membrane proteins by brain capillary endothelial cells in a primary murine model of the blood-brain barrier. In Final Programme, 19th International Symposium on Signal Transduction at the Blood-Brain Barriers, 14-16 September 2016, Copenhagen, Denmark. University of Copenhagen. 2016. p. 37. P-12