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

Publikation: Bidrag til bog/antologi/rapport/konference proceedingKonferenceabstrakt i proceedingForskningpeer review

Resumé

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.
OriginalsprogEngelsk
TitelFinal Programme, 19th International Symposium on Signal Transduction at the Blood-Brain Barriers, 14-16 September 2016, Copenhagen, Denmark
ForlagUniversity of Copenhagen
Publikationsdato2016
Sider37
ArtikelnummerP-12
StatusUdgivet - 2016
Begivenhed19th International Symposium on Signal Transduction at the Blood-Brain Barriers - København, Danmark
Varighed: 14 sep. 201616 sep. 2016

Konference

Konference19th International Symposium on Signal Transduction at the Blood-Brain Barriers
LandDanmark
ByKøbenhavn
Periode14/09/201616/09/2016

Citer dette

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. I Final Programme, 19th International Symposium on Signal Transduction at the Blood-Brain Barriers, 14-16 September 2016, Copenhagen, Denmark (s. 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. s. 37
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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.",
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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. i Final Programme, 19th International Symposium on Signal Transduction at the Blood-Brain Barriers, 14-16 September 2016, Copenhagen, Denmark., P-12, University of Copenhagen, s. 37, 19th International Symposium on Signal Transduction at the Blood-Brain Barriers, København, Danmark, 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. s. 37 P-12.

Publikation: Bidrag til bog/antologi/rapport/konference proceedingKonferenceabstrakt i proceedingForskningpeer 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. I Final Programme, 19th International Symposium on Signal Transduction at the Blood-Brain Barriers, 14-16 September 2016, Copenhagen, Denmark. University of Copenhagen. 2016. s. 37. P-12