Comparison of a Rat Primary Cell-Based Blood-Brain Barrier Model With Epithelial and Brain Endothelial Cell Lines: Gene Expression and Drug Transport
Cell culture-based blood-brain barrier (BBB) models are useful tools for screening of CNS drug candidates. Cell sources for BBB models include primary brain endothelial cells or immortalized brain endothelial cell lines. Despite their well-known differences, epithelial cell lines are also used as su...
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2018-05-01
|
| Series: | Frontiers in Molecular Neuroscience |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/article/10.3389/fnmol.2018.00166/full |
| _version_ | 1818016522353770496 |
|---|---|
| author | Szilvia Veszelka András Tóth András Tóth Fruzsina R. Walter Andrea E. Tóth Ilona Gróf Ilona Gróf Mária Mészáros Mária Mészáros Alexandra Bocsik Éva Hellinger Monika Vastag Gábor Rákhely Gábor Rákhely Mária A. Deli |
| author_facet | Szilvia Veszelka András Tóth András Tóth Fruzsina R. Walter Andrea E. Tóth Ilona Gróf Ilona Gróf Mária Mészáros Mária Mészáros Alexandra Bocsik Éva Hellinger Monika Vastag Gábor Rákhely Gábor Rákhely Mária A. Deli |
| author_sort | Szilvia Veszelka |
| collection | DOAJ |
| description | Cell culture-based blood-brain barrier (BBB) models are useful tools for screening of CNS drug candidates. Cell sources for BBB models include primary brain endothelial cells or immortalized brain endothelial cell lines. Despite their well-known differences, epithelial cell lines are also used as surrogate models for testing neuropharmaceuticals. The aim of the present study was to compare the expression of selected BBB related genes including tight junction proteins, solute carriers (SLC), ABC transporters, metabolic enzymes and to describe the paracellular properties of nine different culture models. To establish a primary BBB model rat brain capillary endothelial cells were co-cultured with rat pericytes and astrocytes (EPA). As other BBB and surrogate models four brain endothelial cells lines, rat GP8 and RBE4 cells, and human hCMEC/D3 cells with or without lithium treatment (D3 and D3L), and four epithelial cell lines, native human intestinal Caco-2 and high P-glycoprotein expressing vinblastine-selected VB-Caco-2 cells, native MDCK and MDR1 transfected MDCK canine kidney cells were used. To test transporter functionality, the permeability of 12 molecules, glucopyranose, valproate, baclofen, gabapentin, probenecid, salicylate, rosuvastatin, pravastatin, atorvastatin, tacrine, donepezil, was also measured in the EPA and epithelial models. Among the junctional protein genes, the expression level of occludin was high in all models except the GP8 and RBE4 cells, and each model expressed a unique claudin pattern. Major BBB efflux (P-glycoprotein or ABCB1) and influx transporters (GLUT-1, LAT-1) were present in all models at mRNA levels. The transcript of BCRP (ABCG2) was not expressed in MDCK, GP8 and RBE4 cells. The absence of gene expression of important BBB efflux and influx transporters BCRP, MRP6, -9, MCT6, -8, PHT2, OATPs in one or both types of epithelial models suggests that Caco-2 or MDCK models are not suitable to test drug candidates which are substrates of these transporters. Brain endothelial cell lines GP8, RBE4, D3 and D3L did not form a restrictive paracellular barrier necessary for screening small molecular weight pharmacons. Therefore, among the tested culture models, the primary cell-based EPA model is suitable for the functional analysis of the BBB. |
| first_indexed | 2024-04-14T07:14:44Z |
| format | Article |
| id | doaj.art-6f2cfed27e264b30bf92342aba80f5ef |
| institution | Directory Open Access Journal |
| issn | 1662-5099 |
| language | English |
| last_indexed | 2024-04-14T07:14:44Z |
| publishDate | 2018-05-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Molecular Neuroscience |
| spelling | doaj.art-6f2cfed27e264b30bf92342aba80f5ef2022-12-22T02:06:20ZengFrontiers Media S.A.Frontiers in Molecular Neuroscience1662-50992018-05-011110.3389/fnmol.2018.00166344242Comparison of a Rat Primary Cell-Based Blood-Brain Barrier Model With Epithelial and Brain Endothelial Cell Lines: Gene Expression and Drug TransportSzilvia Veszelka0András Tóth1András Tóth2Fruzsina R. Walter3Andrea E. Tóth4Ilona Gróf5Ilona Gróf6Mária Mészáros7Mária Mészáros8Alexandra Bocsik9Éva Hellinger10Monika Vastag11Gábor Rákhely12Gábor Rákhely13Mária A. Deli14Biological Barriers Research Group, Institute of Biophysics, Biological Research Centre, Hungarian Academy of Sciences, Szeged, HungaryBiological Barriers Research Group, Institute of Biophysics, Biological Research Centre, Hungarian Academy of Sciences, Szeged, HungaryDepartment of Biotechnology, Faculty of Science and Informatics, University of Szeged, Szeged, HungaryBiological Barriers Research Group, Institute of Biophysics, Biological Research Centre, Hungarian Academy of Sciences, Szeged, HungaryBiological Barriers Research Group, Institute of Biophysics, Biological Research Centre, Hungarian Academy of Sciences, Szeged, HungaryBiological Barriers Research Group, Institute of Biophysics, Biological Research Centre, Hungarian Academy of Sciences, Szeged, HungaryDoctoral School in Biology, Faculty of Science and Informatics, University of Szeged, Szeged, HungaryBiological Barriers Research Group, Institute of Biophysics, Biological Research Centre, Hungarian Academy of Sciences, Szeged, HungaryDoctoral School in Theoretical Medicine, Faculty of Medicine, University of Szeged, Szeged, HungaryBiological Barriers Research Group, Institute of Biophysics, Biological Research Centre, Hungarian Academy of Sciences, Szeged, HungaryIn Vitro Metabolism Research, Division of Pharmacology and Drug Safety, Gedeon Richter Plc., Budapest, HungaryIn Vitro Metabolism Research, Division of Pharmacology and Drug Safety, Gedeon Richter Plc., Budapest, HungaryBiological Barriers Research Group, Institute of Biophysics, Biological Research Centre, Hungarian Academy of Sciences, Szeged, HungaryDepartment of Biotechnology, Faculty of Science and Informatics, University of Szeged, Szeged, HungaryBiological Barriers Research Group, Institute of Biophysics, Biological Research Centre, Hungarian Academy of Sciences, Szeged, HungaryCell culture-based blood-brain barrier (BBB) models are useful tools for screening of CNS drug candidates. Cell sources for BBB models include primary brain endothelial cells or immortalized brain endothelial cell lines. Despite their well-known differences, epithelial cell lines are also used as surrogate models for testing neuropharmaceuticals. The aim of the present study was to compare the expression of selected BBB related genes including tight junction proteins, solute carriers (SLC), ABC transporters, metabolic enzymes and to describe the paracellular properties of nine different culture models. To establish a primary BBB model rat brain capillary endothelial cells were co-cultured with rat pericytes and astrocytes (EPA). As other BBB and surrogate models four brain endothelial cells lines, rat GP8 and RBE4 cells, and human hCMEC/D3 cells with or without lithium treatment (D3 and D3L), and four epithelial cell lines, native human intestinal Caco-2 and high P-glycoprotein expressing vinblastine-selected VB-Caco-2 cells, native MDCK and MDR1 transfected MDCK canine kidney cells were used. To test transporter functionality, the permeability of 12 molecules, glucopyranose, valproate, baclofen, gabapentin, probenecid, salicylate, rosuvastatin, pravastatin, atorvastatin, tacrine, donepezil, was also measured in the EPA and epithelial models. Among the junctional protein genes, the expression level of occludin was high in all models except the GP8 and RBE4 cells, and each model expressed a unique claudin pattern. Major BBB efflux (P-glycoprotein or ABCB1) and influx transporters (GLUT-1, LAT-1) were present in all models at mRNA levels. The transcript of BCRP (ABCG2) was not expressed in MDCK, GP8 and RBE4 cells. The absence of gene expression of important BBB efflux and influx transporters BCRP, MRP6, -9, MCT6, -8, PHT2, OATPs in one or both types of epithelial models suggests that Caco-2 or MDCK models are not suitable to test drug candidates which are substrates of these transporters. Brain endothelial cell lines GP8, RBE4, D3 and D3L did not form a restrictive paracellular barrier necessary for screening small molecular weight pharmacons. Therefore, among the tested culture models, the primary cell-based EPA model is suitable for the functional analysis of the BBB.https://www.frontiersin.org/article/10.3389/fnmol.2018.00166/fullblood-brain barrierbrain endothelial cellsCaco-2MDCKRBE4hCMEC/D3 |
| spellingShingle | Szilvia Veszelka András Tóth András Tóth Fruzsina R. Walter Andrea E. Tóth Ilona Gróf Ilona Gróf Mária Mészáros Mária Mészáros Alexandra Bocsik Éva Hellinger Monika Vastag Gábor Rákhely Gábor Rákhely Mária A. Deli Comparison of a Rat Primary Cell-Based Blood-Brain Barrier Model With Epithelial and Brain Endothelial Cell Lines: Gene Expression and Drug Transport Frontiers in Molecular Neuroscience blood-brain barrier brain endothelial cells Caco-2 MDCK RBE4 hCMEC/D3 |
| title | Comparison of a Rat Primary Cell-Based Blood-Brain Barrier Model With Epithelial and Brain Endothelial Cell Lines: Gene Expression and Drug Transport |
| title_full | Comparison of a Rat Primary Cell-Based Blood-Brain Barrier Model With Epithelial and Brain Endothelial Cell Lines: Gene Expression and Drug Transport |
| title_fullStr | Comparison of a Rat Primary Cell-Based Blood-Brain Barrier Model With Epithelial and Brain Endothelial Cell Lines: Gene Expression and Drug Transport |
| title_full_unstemmed | Comparison of a Rat Primary Cell-Based Blood-Brain Barrier Model With Epithelial and Brain Endothelial Cell Lines: Gene Expression and Drug Transport |
| title_short | Comparison of a Rat Primary Cell-Based Blood-Brain Barrier Model With Epithelial and Brain Endothelial Cell Lines: Gene Expression and Drug Transport |
| title_sort | comparison of a rat primary cell based blood brain barrier model with epithelial and brain endothelial cell lines gene expression and drug transport |
| topic | blood-brain barrier brain endothelial cells Caco-2 MDCK RBE4 hCMEC/D3 |
| url | https://www.frontiersin.org/article/10.3389/fnmol.2018.00166/full |
| work_keys_str_mv | AT szilviaveszelka comparisonofaratprimarycellbasedbloodbrainbarriermodelwithepithelialandbrainendothelialcelllinesgeneexpressionanddrugtransport AT andrastoth comparisonofaratprimarycellbasedbloodbrainbarriermodelwithepithelialandbrainendothelialcelllinesgeneexpressionanddrugtransport AT andrastoth comparisonofaratprimarycellbasedbloodbrainbarriermodelwithepithelialandbrainendothelialcelllinesgeneexpressionanddrugtransport AT fruzsinarwalter comparisonofaratprimarycellbasedbloodbrainbarriermodelwithepithelialandbrainendothelialcelllinesgeneexpressionanddrugtransport AT andreaetoth comparisonofaratprimarycellbasedbloodbrainbarriermodelwithepithelialandbrainendothelialcelllinesgeneexpressionanddrugtransport AT ilonagrof comparisonofaratprimarycellbasedbloodbrainbarriermodelwithepithelialandbrainendothelialcelllinesgeneexpressionanddrugtransport AT ilonagrof comparisonofaratprimarycellbasedbloodbrainbarriermodelwithepithelialandbrainendothelialcelllinesgeneexpressionanddrugtransport AT mariameszaros comparisonofaratprimarycellbasedbloodbrainbarriermodelwithepithelialandbrainendothelialcelllinesgeneexpressionanddrugtransport AT mariameszaros comparisonofaratprimarycellbasedbloodbrainbarriermodelwithepithelialandbrainendothelialcelllinesgeneexpressionanddrugtransport AT alexandrabocsik comparisonofaratprimarycellbasedbloodbrainbarriermodelwithepithelialandbrainendothelialcelllinesgeneexpressionanddrugtransport AT evahellinger comparisonofaratprimarycellbasedbloodbrainbarriermodelwithepithelialandbrainendothelialcelllinesgeneexpressionanddrugtransport AT monikavastag comparisonofaratprimarycellbasedbloodbrainbarriermodelwithepithelialandbrainendothelialcelllinesgeneexpressionanddrugtransport AT gaborrakhely comparisonofaratprimarycellbasedbloodbrainbarriermodelwithepithelialandbrainendothelialcelllinesgeneexpressionanddrugtransport AT gaborrakhely comparisonofaratprimarycellbasedbloodbrainbarriermodelwithepithelialandbrainendothelialcelllinesgeneexpressionanddrugtransport AT mariaadeli comparisonofaratprimarycellbasedbloodbrainbarriermodelwithepithelialandbrainendothelialcelllinesgeneexpressionanddrugtransport |