Improved Method for the Establishment of an &lt;em&gt;In Vitro&lt;/em&gt; Blood-Brain Barrier Model Based on Porcine Brain Endothelial Cells

Nielsen, Simone S. E.; Siupka, Piotr; Georgian, Ana Ruxandra; Preston, Jane E.; Tóth, Anna Zsófia; Yusof, Siti R.; Abbott, N. Joan; Nielsen, Mogens

doi:10.3791/56277-v

Cited by 4 publications

(4 citation statements)

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“…As shown in Figure 7 A–C , the protein and mRNA expression of ZO‐1, Occludin, and Claudin‐5 was significantly decreased after α ‐Syn PFFs administration. Furthermore, as shown in Figure 7D , α ‐Syn PFFs administration reduced the TEER value (on day 3 to day 7) in both the coculture system of BMVECs and astrocytes (in vitro BBB model [ 28 ] ) and the BMVEC monoculture system.…”

Section: Resultsmentioning

confidence: 99%

“…Two systems, including a noncontact coculture system of primary BMVECs and primary astrocytes (an in vitro BBB model [ 28 ] ) and a separate primary BMVEC culture system, were used to detect epithelial cell barrier integrity and permeability. For the coculture system, a transwell chamber with a pore size of 0.4 µm was used to separate BMVECs and astrocytes.…”

Section: Methodsmentioning

confidence: 99%

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Cerebral Microvascular Injury Induced by Lag3‐Dependent α‐Synuclein Fibril Endocytosis Exacerbates Cognitive Impairment in a Mouse Model of α‐Synucleinopathies

et al. 2023

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The pathological accumulation of α‐synuclein (α‐Syn) and the transmission of misfolded α‐Syn underlie α‐synucleinopathies. Increased plasma α‐Syn levels are associated with cognitive impairment in Parkinson's disease, multiple system atrophy, and dementia with Lewy bodies, but it is still unknown whether the cognitive deficits in α‐synucleinopathies have a common vascular pathological origin. Here, it is reported that combined injection of α‐Syn preformed fibrils (PFFs) in the unilateral substantia nigra pars compacta, hippocampus, and cerebral cortex results in impaired spatial learning and memory abilities at 6 months post‐injection and that this cognitive decline is related to cerebral microvascular injury. Moreover, insoluble α‐Syn inclusions are found to form in primary mouse brain microvascular endothelial cells (BMVECs) through lymphocyte‐activation gene 3 (Lag3)‐dependent α‐Syn PFFs endocytosis, causing poly(ADP‐ribose)‐driven cell death and reducing the expression of tight junction proteins in BMVECs. Knockout of Lag3 in vitro prevents α‐Syn PFFs from entering BMVECs, thereby reducing the abovementioned response induced by α‐Syn PFFs. Deletion of endothelial cell‐specific Lag3 in vivo reverses the negative effects of α‐Syn PFFs on cerebral microvessels and cognitive function. In short, this study reveals the effectiveness of targeting Lag3 to block the spread of α‐Syn fibrils to endothelial cells in order to improve cognition.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Cerebral Microvascular Injury Induced by Lag3‐Dependent α‐Synuclein Fibril Endocytosis Exacerbates Cognitive Impairment in a Mouse Model of α‐Synucleinopathies

et al. 2023

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“…In practice, multi‐cell co‐culture models are mostly used. The multicell co‐culture model contains two or more types of cells, and the culture methods are subdivided into contact co‐culture and non‐contact co‐culture (Nielsen et al., 2017; Sivandzade & Cucullo, 2018). Co‐culture of endothelial cells with several kinds of other cells in different ways (Figure 2) mimics the structure and function of the in vivo BBB even better (Hatherell et al., 2011; Stone et al., 2019).…”

Section: Transwell Insert‐based Modelsmentioning

confidence: 99%

Progresses in the establishment, evaluation, and application of in vitro blood–brain barrier models

Yin,

Wang

2024

J of Neuroscience Research

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The blood–brain barrier (BBB) is a barrier between the circulatory system and the central nervous system (CNS), contributing to CNS protection and maintaining the brain homeostasis. Establishment of in vitro BBB models that are closer to the microenvironment of the human brain is helpful for evaluating the potential and efficiency of a drug penetrating BBB and thus the clinical application value of the drug. The in vitro BBB models not only provide great convenience for screening new drugs that can access to CNS but also help people to have a deeper study on the mechanism of substances entering and leaving the brain, which makes people have greater opportunities in the treatment of CNS diseases. Up to now, although much effort has been paid to the researches on the in vitro BBB models and many progresses have been achieved, no unified method has been described for establishing a BBB model and there is much work to do and many challenges to be faced with in the future. This review summarizes the research progresses in the establishment, evaluation, and application of in vitro BBB models.

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“…Porcine brain endothelial cells (PBEC, BBB cells) were isolated and cultured as previously published [45]. Cells were seeded onto collagen and fibronectin coated polycarbonate Transwell filters (Corning, 1.12cm 2 filter surface area, 0.4 μm pore size), at 10 × 10 4 cells per filter in 12 well plates, and grown above Poly-L-Lysine-coated wells containing confluent primary rat astrocytes.…”

Section: Blood Brain Barrier Cell Culturementioning

confidence: 99%

Glucosamine-NISV delivers antibody across the blood-brain barrier: Optimization for treatment of encephalitic viruses

Woods

O’Brien

Butcher

et al. 2020

Journal of Controlled Release

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Improved Method for the Establishment of an <em>In Vitro</em> Blood-Brain Barrier Model Based on Porcine Brain Endothelial Cells

Cited by 4 publications

References 0 publications

Cerebral Microvascular Injury Induced by Lag3‐Dependent α‐Synuclein Fibril Endocytosis Exacerbates Cognitive Impairment in a Mouse Model of α‐Synucleinopathies

Cerebral Microvascular Injury Induced by Lag3‐Dependent α‐Synuclein Fibril Endocytosis Exacerbates Cognitive Impairment in a Mouse Model of α‐Synucleinopathies

Progresses in the establishment, evaluation, and application of in vitro blood–brain barrier models

Glucosamine-NISV delivers antibody across the blood-brain barrier: Optimization for treatment of encephalitic viruses

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