Cargando…
Establishment of a simplified in vitro porcine blood–brain barrier model with high transendothelial electrical resistance
Good in vitro blood–brain barrier (BBB) models that mimic the in vivo BBB phenotype are essential for studies on BBB functionality and for initial screening in drug discovery programmes, as many potential therapeutic drug candidates have poor BBB permeation. Difficulties associated with the availabi...
Autores principales: | , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier/North-Holland Biomedical Press
2013
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3694297/ https://www.ncbi.nlm.nih.gov/pubmed/22789905 http://dx.doi.org/10.1016/j.brainres.2012.06.057 |
_version_ | 1782274844464775168 |
---|---|
author | Patabendige, Adjanie Skinner, Robert A. Abbott, N. Joan |
author_facet | Patabendige, Adjanie Skinner, Robert A. Abbott, N. Joan |
author_sort | Patabendige, Adjanie |
collection | PubMed |
description | Good in vitro blood–brain barrier (BBB) models that mimic the in vivo BBB phenotype are essential for studies on BBB functionality and for initial screening in drug discovery programmes, as many potential therapeutic drug candidates have poor BBB permeation. Difficulties associated with the availability of human brain tissue, coupled with the time and cost associated with using animals for this kind of research have led to the development of non-human cell culture models. However, most BBB models display a low transendothelial electrical resistance (TEER), which is a measure of the tightness of the BBB. To address these issues we have established and optimised a robust, simple to use in vitro BBB model using porcine brain endothelial cells (PBECs). The PBEC model gives high TEER without the need for co-culture with astrocytes (up to 1300 Ω cm(2) with a mean TEER of ∼800 Ω cm(2)) with well organised tight junctions as shown by immunostaining for occludin and claudin-5. Functional assays confirmed the presence of high levels of alkaline phosphatase (ALP), and presence of the efflux transporter, P-glycoprotein (P-gp, ABCB1). Presence of the breast cancer resistance protein (BCRP, ABCG2) was confirmed by TaqMan real-time RT-PCR assay. Real-time RT-PCR assays for BCRP, occludin and claudin-5 demonstrated no significant differences between batches of PBECs, and also between primary and passage 1 PBECs. A permeability screen of 10 compounds demonstrated the usefulness of the model as a tool for drug permeability studies. Qualitative and quantitative results from this study confirm that this in vitro porcine BBB model is reliable and robust; it is also simpler to generate than most other BBB models. This article is part of a Special Issue entitled Electrical Synapses. |
format | Online Article Text |
id | pubmed-3694297 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Elsevier/North-Holland Biomedical Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-36942972013-07-12 Establishment of a simplified in vitro porcine blood–brain barrier model with high transendothelial electrical resistance Patabendige, Adjanie Skinner, Robert A. Abbott, N. Joan Brain Res Research Report Good in vitro blood–brain barrier (BBB) models that mimic the in vivo BBB phenotype are essential for studies on BBB functionality and for initial screening in drug discovery programmes, as many potential therapeutic drug candidates have poor BBB permeation. Difficulties associated with the availability of human brain tissue, coupled with the time and cost associated with using animals for this kind of research have led to the development of non-human cell culture models. However, most BBB models display a low transendothelial electrical resistance (TEER), which is a measure of the tightness of the BBB. To address these issues we have established and optimised a robust, simple to use in vitro BBB model using porcine brain endothelial cells (PBECs). The PBEC model gives high TEER without the need for co-culture with astrocytes (up to 1300 Ω cm(2) with a mean TEER of ∼800 Ω cm(2)) with well organised tight junctions as shown by immunostaining for occludin and claudin-5. Functional assays confirmed the presence of high levels of alkaline phosphatase (ALP), and presence of the efflux transporter, P-glycoprotein (P-gp, ABCB1). Presence of the breast cancer resistance protein (BCRP, ABCG2) was confirmed by TaqMan real-time RT-PCR assay. Real-time RT-PCR assays for BCRP, occludin and claudin-5 demonstrated no significant differences between batches of PBECs, and also between primary and passage 1 PBECs. A permeability screen of 10 compounds demonstrated the usefulness of the model as a tool for drug permeability studies. Qualitative and quantitative results from this study confirm that this in vitro porcine BBB model is reliable and robust; it is also simpler to generate than most other BBB models. This article is part of a Special Issue entitled Electrical Synapses. Elsevier/North-Holland Biomedical Press 2013-07-12 /pmc/articles/PMC3694297/ /pubmed/22789905 http://dx.doi.org/10.1016/j.brainres.2012.06.057 Text en © 2013 Elsevier B.V. https://creativecommons.org/licenses/by/3.0/ Open Access under CC BY 3.0 (https://creativecommons.org/licenses/by/3.0/) license |
spellingShingle | Research Report Patabendige, Adjanie Skinner, Robert A. Abbott, N. Joan Establishment of a simplified in vitro porcine blood–brain barrier model with high transendothelial electrical resistance |
title | Establishment of a simplified in vitro porcine blood–brain barrier model with high transendothelial electrical resistance |
title_full | Establishment of a simplified in vitro porcine blood–brain barrier model with high transendothelial electrical resistance |
title_fullStr | Establishment of a simplified in vitro porcine blood–brain barrier model with high transendothelial electrical resistance |
title_full_unstemmed | Establishment of a simplified in vitro porcine blood–brain barrier model with high transendothelial electrical resistance |
title_short | Establishment of a simplified in vitro porcine blood–brain barrier model with high transendothelial electrical resistance |
title_sort | establishment of a simplified in vitro porcine blood–brain barrier model with high transendothelial electrical resistance |
topic | Research Report |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3694297/ https://www.ncbi.nlm.nih.gov/pubmed/22789905 http://dx.doi.org/10.1016/j.brainres.2012.06.057 |
work_keys_str_mv | AT patabendigeadjanie establishmentofasimplifiedinvitroporcinebloodbrainbarriermodelwithhightransendothelialelectricalresistance AT skinnerroberta establishmentofasimplifiedinvitroporcinebloodbrainbarriermodelwithhightransendothelialelectricalresistance AT abbottnjoan establishmentofasimplifiedinvitroporcinebloodbrainbarriermodelwithhightransendothelialelectricalresistance |