Quantitative Targeted Absolute Proteomics for Better Characterization of an In Vitro Human Blood–Brain Barrier Model Derived from Hematopoietic Stem Cells

We previously developed an in vitro model of the human blood–brain barrier (BBB) based on the use of endothelial cells derived from CD34+-hematopoietic stem cells and cultured with brain pericytes. The purpose of the present study was to provide information on the protein expression levels of the tr...

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Vydáno v:	Cells (Basel, Switzerland) Ročník 11; číslo 24; s. 3963
Hlavní autoři:	Dehouck, Marie-Pierre, Tachikawa, Masanori, Hoshi, Yutaro, Omori, Kotaro, Maurage, Claude-Alain, Strecker, Guillaume, Dehouck, Lucie, Boucau, Marie-Christine, Uchida, Yasuo, Gosselet, Fabien, Terasaki, Tetsuya, Karamanos, Yannis
Médium:	Journal Article
Jazyk:	angličtina
Vydáno:	Switzerland MDPI AG 08.12.2022 MDPI
Témata:	Analysis ATP Binding Cassette Transporter, Subfamily G, Member 2 - metabolism Biochemistry, Molecular Biology Blood & organ donations Blood vessels Blood-brain barrier Blood-Brain Barrier - metabolism Brain brain microvessels Caveolin Caveolin-1 CD147 antigen CD34 antigen Chromatography Chromatography, Liquid Claudin-5 - metabolism Connexin 43 Endothelial cells Endothelial Cells - metabolism Genomics Hematopoietic stem cells Hematopoietic Stem Cells - metabolism human brain-like endothelial cells human in vitro models Humans LC-MS/MS-based protein quantification Life Sciences Liquid chromatography Mass spectrometry Mass spectroscopy MDR1 protein Membrane Transport Proteins - metabolism Neoplasm Proteins - metabolism Neurobiology Neurons and Cognition Pericytes Permeability Protein expression Protein folding Proteins Proteomics Proteomics - methods Stem cells Tandem Mass Spectrometry - methods Transplantation transporter France United States > US blood–brain barrier human in vitro models human brain-like endothelial cells LC-MS/MS-based protein quantification brain microvessels transporter LC-MS/MS-based protein quantiﬁcation human brain-like endothelial cell
ISSN:	2073-4409, 2073-4409
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Popis
Shrnutí:	We previously developed an in vitro model of the human blood–brain barrier (BBB) based on the use of endothelial cells derived from CD34+-hematopoietic stem cells and cultured with brain pericytes. The purpose of the present study was to provide information on the protein expression levels of the transporters, receptors, tight junction/adherence junction molecules, and transporter-associated molecules of human brain-like endothelial cells (hBLECs). The absolute protein expression levels were determined by liquid chromatography–mass spectrometry-based quantitative targeted absolute proteomics and compared with those from human brain microvessels (hBMVs). The protein levels of CD144, CD147, MRP4, Annexin A6 and caveolin-1 showed more than 3-fold abundance in hBLECs, those of MCT1, Connexin 43, TfR1, and claudin-5 showed less than 3-fold differences, and the protein levels of other drug efflux transporters and nutrient transporters were less represented in hBLECs than in hBMVs. It is noteworthy that BCRP was more expressed than MDR1 in hBLECs, as this was the case for hBMVs. These results suggest that transports mediated by MCT1, TfR1, and claudin-5-related tight junction function reflect the in vivo BBB situation. The present study provided a better characterization of hBLECs and clarified the equivalence of the transport characteristics between in vitro BBB models and in vivo BBB models using LC-MS/MS-based protein quantification.
Bibliografie:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 14 content type line 23 These authors contributed equally to this work.
ISSN:	2073-4409 2073-4409
DOI:	10.3390/cells11243963