A three-dimensional model of the human blood-brain barrier to analyse the transport of nanoparticles and astrocyte/endothelial interactions [version 2; referees: 2 approved]
The aim of this study was to develop a three-dimensional (3D) model of the human blood-brain barrier in vitro, which mimics the cellular architecture of the CNS and could be used to analyse the delivery of nanoparticles to cells of the CNS. The model includes human astrocytes set in a collagen gel,...
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F1000 Research Ltd
2016-01-01
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| Online Access: | http://f1000research.com/articles/4-1279/v2 |
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| author | Peddagangannagari Sreekanthreddy Radka Gromnicova Heather Davies James Phillips Ignacio A. Romero David Male |
| author_facet | Peddagangannagari Sreekanthreddy Radka Gromnicova Heather Davies James Phillips Ignacio A. Romero David Male |
| author_sort | Peddagangannagari Sreekanthreddy |
| collection | DOAJ |
| description | The aim of this study was to develop a three-dimensional (3D) model of the human blood-brain barrier in vitro, which mimics the cellular architecture of the CNS and could be used to analyse the delivery of nanoparticles to cells of the CNS. The model includes human astrocytes set in a collagen gel, which is overlaid by a monolayer of human brain endothelium (hCMEC/D3 cell line). The model was characterised by transmission electron microscopy (TEM), immunofluorescence microscopy and flow cytometry. A collagenase digestion method could recover the two cell types separately at 92-96% purity. Astrocytes grown in the gel matrix do not divide and they have reduced expression of aquaporin-4 and the endothelin receptor, type B compared to two-dimensional cultures, but maintain their expression of glial fibrillary acidic protein. The effects of conditioned media from these astrocytes on the barrier phenotype of the endothelium was compared with media from astrocytes grown conventionally on a two-dimensional (2D) substratum. Both induce the expression of tight junction proteins zonula occludens-1 and claudin-5 in hCMEC/D3 cells, but there was no difference between the induced expression levels by the two media. The model has been used to assess the transport of glucose-coated 4nm gold nanoparticles and for leukocyte migration. TEM was used to trace and quantitate the movement of the nanoparticles across the endothelium and into the astrocytes. This blood-brain barrier model is very suitable for assessing delivery of nanoparticles and larger biomolecules to cells of the CNS, following transport across the endothelium. |
| first_indexed | 2024-12-22T20:46:24Z |
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| institution | Directory Open Access Journal |
| issn | 2046-1402 |
| language | English |
| last_indexed | 2024-12-22T20:46:24Z |
| publishDate | 2016-01-01 |
| publisher | F1000 Research Ltd |
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| series | F1000Research |
| spelling | doaj.art-8e1e5c0c576b47e6a9e60ca8d03859ad2022-12-21T18:13:12ZengF1000 Research LtdF1000Research2046-14022016-01-01410.12688/f1000research.7142.28385A three-dimensional model of the human blood-brain barrier to analyse the transport of nanoparticles and astrocyte/endothelial interactions [version 2; referees: 2 approved]Peddagangannagari Sreekanthreddy0Radka Gromnicova1Heather Davies2James Phillips3Ignacio A. Romero4David Male5Department of Life, Health and Chemical Sciences, The Open University, Milton Keynes, MK7 6AA, UKDepartment of Life, Health and Chemical Sciences, The Open University, Milton Keynes, MK7 6AA, UKDepartment of Life, Health and Chemical Sciences, The Open University, Milton Keynes, MK7 6AA, UKDepartment of Life, Health and Chemical Sciences, The Open University, Milton Keynes, MK7 6AA, UKDepartment of Life, Health and Chemical Sciences, The Open University, Milton Keynes, MK7 6AA, UKDepartment of Life, Health and Chemical Sciences, The Open University, Milton Keynes, MK7 6AA, UKThe aim of this study was to develop a three-dimensional (3D) model of the human blood-brain barrier in vitro, which mimics the cellular architecture of the CNS and could be used to analyse the delivery of nanoparticles to cells of the CNS. The model includes human astrocytes set in a collagen gel, which is overlaid by a monolayer of human brain endothelium (hCMEC/D3 cell line). The model was characterised by transmission electron microscopy (TEM), immunofluorescence microscopy and flow cytometry. A collagenase digestion method could recover the two cell types separately at 92-96% purity. Astrocytes grown in the gel matrix do not divide and they have reduced expression of aquaporin-4 and the endothelin receptor, type B compared to two-dimensional cultures, but maintain their expression of glial fibrillary acidic protein. The effects of conditioned media from these astrocytes on the barrier phenotype of the endothelium was compared with media from astrocytes grown conventionally on a two-dimensional (2D) substratum. Both induce the expression of tight junction proteins zonula occludens-1 and claudin-5 in hCMEC/D3 cells, but there was no difference between the induced expression levels by the two media. The model has been used to assess the transport of glucose-coated 4nm gold nanoparticles and for leukocyte migration. TEM was used to trace and quantitate the movement of the nanoparticles across the endothelium and into the astrocytes. This blood-brain barrier model is very suitable for assessing delivery of nanoparticles and larger biomolecules to cells of the CNS, following transport across the endothelium.http://f1000research.com/articles/4-1279/v2Cerebrovascular DiseaseExperimental Biophysical MethodsTheory & Simulation |
| spellingShingle | Peddagangannagari Sreekanthreddy Radka Gromnicova Heather Davies James Phillips Ignacio A. Romero David Male A three-dimensional model of the human blood-brain barrier to analyse the transport of nanoparticles and astrocyte/endothelial interactions [version 2; referees: 2 approved] F1000Research Cerebrovascular Disease Experimental Biophysical Methods Theory & Simulation |
| title | A three-dimensional model of the human blood-brain barrier to analyse the transport of nanoparticles and astrocyte/endothelial interactions [version 2; referees: 2 approved] |
| title_full | A three-dimensional model of the human blood-brain barrier to analyse the transport of nanoparticles and astrocyte/endothelial interactions [version 2; referees: 2 approved] |
| title_fullStr | A three-dimensional model of the human blood-brain barrier to analyse the transport of nanoparticles and astrocyte/endothelial interactions [version 2; referees: 2 approved] |
| title_full_unstemmed | A three-dimensional model of the human blood-brain barrier to analyse the transport of nanoparticles and astrocyte/endothelial interactions [version 2; referees: 2 approved] |
| title_short | A three-dimensional model of the human blood-brain barrier to analyse the transport of nanoparticles and astrocyte/endothelial interactions [version 2; referees: 2 approved] |
| title_sort | three dimensional model of the human blood brain barrier to analyse the transport of nanoparticles and astrocyte endothelial interactions version 2 referees 2 approved |
| topic | Cerebrovascular Disease Experimental Biophysical Methods Theory & Simulation |
| url | http://f1000research.com/articles/4-1279/v2 |
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