ArticleJournal of visualized experiments : JoVE2017
Improved Method for the Establishment of an In Vitro Blood-Brain Barrier Model Based on Porcine Brain Endothelial Cells.
Article in Journal of visualized experiments : JoVE, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed, 30 citations in OpenAlex.
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- Ex vivo nanoscale abluminal mapping of putative cargo receptors at the blood-brain barrier of expanded brain capillaries.Fluids and barriers of the CNS · 2024Article
- Cerebral Microvascular Injury Induced by Lag3-Dependent α-Synuclein Fibril Endocytosis Exacerbates Cognitive Impairment in a Mouse Model of α-Synucleinopathies.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2023Article
- Review
- Apicobasal transferrin receptor localization and trafficking in brain capillary endothelial cells.Fluids and barriers of the CNS · 2023Article
- Polarized α-synuclein trafficking and transcytosis across brain endothelial cells via Rab7-decorated carriers.Fluids and barriers of the CNS · 2022Article
- Advances in Non-Animal Testing Approaches towards Accelerated Clinical Translation of Novel Nanotheranostic Therapeutics for Central Nervous System Disorders.Nanomaterials (Basel, Switzerland) · 2021Review
- Basigin Antibodies with Capacity for Drug Delivery Across Brain Endothelial Cells.Molecular neurobiology · 2021Article
- Transport of a Peptide from Bovine αNutrients · 2020Article
- CoQJournal of clinical medicine · 2020Article
- Characterization of basigin monoclonal antibodies for receptor-mediated drug delivery to the brain.Scientific reports · 2020Article
- Investigating Maternal Brain Alterations in Preeclampsia: the Need for a Multidisciplinary Effort.Current hypertension reports · 2019Review
- The endo-lysosomal system of bEnd.3 and hCMEC/D3 brain endothelial cells.Fluids and barriers of the CNS · 2019Article
- Analysis of the trafficking system in blood-brain barrier models by high content screening microscopy.Neural regeneration research · 2018Article
- The Endo-Lysosomal System of Brain Endothelial Cells Is Influenced by Astrocytes In Vitro.Molecular neurobiology · 2018Article
Corrections and comments
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Authors and funding
8 authors at 4 institutions in 3 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The aim of this protocol presents an optimized procedure for the purification and cultivation of pBECs and to establish in vitro blood-brain barrier (BBB) models based on pBECs in mono-culture (MC), MC with astrocyte-conditioned medium (ACM), and non-contact co-culture (NCC) with astrocytes of porcine or rat origin. pBECs were isolated and cultured from fragments of capillaries from the brain cortices of domestic pigs 5-6 months old. These fragments were purified by careful removal of meninges, isolation and homogenization of grey matter, filtration, enzymatic digestion, and centrifugation. To further eliminate contaminating cells, the capillary fragments were cultured with puromycin-containing medium. When 60-95% confluent, pBECs growing from the capillary fragments were passaged to permeable membrane filter inserts and established in the models. To increase barrier tightness and BBB characteristic phenotype of pBECs, the cells were treated with the following differentiation factors: membrane permeant 8-CPT-cAMP (here abbreviated cAMP), hydrocortisone, and a phosphodiesterase inhibitor, RO-20-1724 (RO). The procedure was carried out over a period of 9-11 days, and when establishing the NCC model, the astrocytes were cultured 2-8 weeks in advance. Adherence to the described procedures in the protocol has allowed the establishment of endothelial layers with highly restricted paracellular permeability, with the NCC model showing an average transendothelial electrical resistance (TEER) of 1249 ± 80 Ω cm
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.