Recent advances in human iPSC-derived models of the blood–brain barrier

Workman, Michael J.; Svendsen, Clive N.

doi:10.1186/s12987-020-00191-7

Cited by 99 publications

(78 citation statements)

References 87 publications

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“…However, the underlying interspecies differences query the reliability and prediction capacity of most in vitro BBB models for the passage of therapeutic molecules across the human BBB and into the CNS [ 4 , 5 ]. More recently, human BBB models from induced pluripotent stem cell (iPSC)-derived endothelial cells have been developed, but it is still uncertain to what extent they may recapitulate the BBB and predict in vivo exposures [ 6 ].…”

Section: Introductionmentioning

confidence: 99%

Non-Human Primate Blood–Brain Barrier and In Vitro Brain Endothelium: From Transcriptome to the Establishment of a New Model

Chaves

Cegarra

et al. 2020

Pharmaceutics

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The non-human primate (NHP)-brain endothelium constitutes an essential alternative to human in the prediction of molecule trafficking across the blood–brain barrier (BBB). This study presents a comparison between the NHP transcriptome of freshly isolated brain microcapillaries and in vitro-selected brain endothelial cells (BECs), focusing on important BBB features, namely tight junctions, receptors mediating transcytosis (RMT), ABC and SLC transporters, given its relevance as an alternative model for the molecule trafficking prediction across the BBB and identification of new brain-specific transport mechanisms. In vitro BECs conserved most of the BBB key elements for barrier integrity and control of molecular trafficking. The function of RMT via the transferrin receptor (TFRC) was characterized in this NHP-BBB model, where both human transferrin and anti-hTFRC antibody showed increased apical-to-basolateral passage in comparison to control molecules. In parallel, eventual BBB-related regional differences were investigated in seven-day in vitro-selected BECs from five brain structures: brainstem, cerebellum, cortex, hippocampus, and striatum. Our analysis retrieved few differences in the brain endothelium across brain regions, suggesting a rather homogeneous BBB function across the brain parenchyma. The presently established NHP-derived BBB model closely mimics the physiological BBB, thus representing a ready-to-use tool for assessment of the penetration of biotherapeutics into the human CNS.

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Section: Introductionmentioning

confidence: 99%

Non-Human Primate Blood–Brain Barrier and In Vitro Brain Endothelium: From Transcriptome to the Establishment of a New Model

Chaves

Cegarra

et al. 2020

Pharmaceutics

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“…In addition, expression of VE-cadherin, an essential endothelial marker, is not distinctly junctional in some reports [ 6 – 8 ]. There is also concern about the developmental stage and maturation of stem cell-derived cells, including iBMECs [ 9 ]. Shear stress acting on endothelial cells due to the presence of flow is a significant factor that contributes to endothelial differentiation and function [ 10 – 13 ].…”

Section: Introductionmentioning

confidence: 99%

Chemically defined human vascular laminins for biologically relevant culture of hiPSC-derived brain microvascular endothelial cells

Motallebnejad

Azarin

2020

Fluids Barriers CNS

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Background In recent years, differentiation of human induced pluripotent stem cells (hiPSCs) into brain-specific microvascular endothelial cells (iBMECs) has frequently been used to model the blood–brain barrier (BBB). However, there are limitations in the use of iBMECs for in vitro studies, such as transendothelial electrical resistance (TEER) instability, weak junctional expression of VE-cadherin, and lack of proper fluid shear stress response. In vivo, the basement membrane (BM) composition of the BBB evolves throughout development, and laminins become the dominant component of the mature vascular BM. However, laminin isoforms of the endothelial BM have not been used for culture of differentiated iBMECs. The main goal of this study is to investigate the effect of different laminin isoforms of the endothelial BM on iBMEC functionality and to determine whether better recapitulation of the physiological BM in vitro can address the aforementioned limitations of iBMECs. Methods Using a previously reported method, hiPSCs were differentiated into iBMECs. The influence of main laminins of the endothelial BM, LN 411 and LN 511, on iBMEC functionality was studied and compared to a collagen IV and fibronectin mixture (CN IV-FN). Quantitative RT-PCR, immunocytochemistry, and TEER measurement were utilized to assess gene and protein expression and barrier properties of iBMECs on different extracellular matrices. Single-channel microfluidic devices were used to study the effect of shear stress on iBMECs. Results LN 511, but not LN 411, improved iBMEC barrier properties and resulted in more sustained TEER stability. Immunocytochemistry showed improved junctional protein expression compared to iBMECs cultured on CN IV-FN. iBMECs cultured on LN 511 showed a reduction of stress fibers, indicating resting endothelial phenotype, whereas gene expression analysis revealed upregulation of multiple genes involved in endothelial activation in iBMECs on CN IV-FN. Finally, culturing iBMECs on LN 511 enhanced physiological responses to shear stress, including morphological changes and enhanced junctional protein association. Conclusion LN 511 improves the functionality and long-term barrier stability of iBMECs. Our findings suggest that incorporation of physiologically relevant LN 511 in iBMEC culture would be beneficial for disease modeling applications and BBB-on-a-chip platforms that accommodate fluid flow.

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“…Recent advancements in stem cell technology have furthermore allowed derivation of human in vitro models of the BBB from stem cell sources including human cord blood-derived stem cells of circulating endothelial progenitors [98] and human induced pluripotent stem cells (hiPSCs; summarized in [99]). hiPSCs derived from one individual opens the entirely novel opportunity to study BBB dysfunction from individual patients as their hiPSCs provide a scalable and renewable source for establishing brain microvascular endothelial cells.…”

Section: Bbb In Vitro Models: Cultured Primary Brain Endothelial Cellmentioning

confidence: 99%

Advancing brain barriers RNA sequencing: guidelines from experimental design to publication

Francisco

Marchetti

Rodríguez‐Lorenzo

et al. 2020

Fluids Barriers CNS

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Background: RNA sequencing (RNA-Seq) in its varied forms has become an indispensable tool for analyzing differential gene expression and thus characterization of specific tissues. Aiming to understand the brain barriers genetic signature, RNA seq has also been introduced in brain barriers research. This has led to availability of both, bulk and single-cell RNA-Seq datasets over the last few years. If appropriately performed, the RNA-Seq studies provide powerful datasets that allow for significant deepening of knowledge on the molecular mechanisms that establish the brain barriers. However, RNA-Seq studies comprise complex workflows that require to consider many options and variables before, during and after the proper sequencing process. Main body: In the current manuscript, we build on the interdisciplinary experience of the European PhD Training Network BtRAIN (https ://www.btrai n-2020.eu/) where bioinformaticians and brain barriers researchers collaborated to analyze and establish RNA-Seq datasets on vertebrate brain barriers. The obstacles BtRAIN has identified in this process have been integrated into the present manuscript. It provides guidelines along the entire workflow of brain barriers RNA-Seq studies starting from the overall experimental design to interpretation of results. Focusing on the vertebrate endothelial blood-brain barrier (BBB) and epithelial blood-cerebrospinal-fluid barrier (BCSFB) of the choroid plexus, we provide a step-by-step description of the workflow, highlighting the decisions to be made at each step of the workflow and explaining the strengths and weaknesses of individual choices made. Finally, we propose recommendations for accurate data interpretation and on the information to be included into a publication to ensure appropriate accessibility of the data and reproducibility of the observations by the scientific community. Conclusion: Next generation transcriptomic profiling of the brain barriers provides a novel resource for understanding the development, function and pathology of these barrier cells, which is essential for understanding CNS homeostasis and disease. Continuous advancement and sophistication of RNA-Seq will require interdisciplinary approaches between brain barrier researchers and bioinformaticians as successfully performed in BtRAIN. The present guidelines are built on the BtRAIN interdisciplinary experience and aim to facilitate collaboration of brain barriers researchers with bioinformaticians to advance RNA-Seq study design in the brain barriers community.

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Recent advances in human iPSC-derived models of the blood–brain barrier

Cited by 99 publications

References 87 publications

Non-Human Primate Blood–Brain Barrier and In Vitro Brain Endothelium: From Transcriptome to the Establishment of a New Model

Non-Human Primate Blood–Brain Barrier and In Vitro Brain Endothelium: From Transcriptome to the Establishment of a New Model

Chemically defined human vascular laminins for biologically relevant culture of hiPSC-derived brain microvascular endothelial cells

Advancing brain barriers RNA sequencing: guidelines from experimental design to publication

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