A perfused human blood–brain barrier on-a-chip for high-throughput assessment of barrier function and antibody transport
Abstract Background Receptor-mediated transcytosis is one of the major routes for drug delivery of large molecules into the brain. The aim of this study was to develop a novel model of the human blood–brain barrier (BBB) in a high-throughput microfluidic device. This model can be used to assess pass...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2018-08-01
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| Series: | Fluids and Barriers of the CNS |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12987-018-0108-3 |
| _version_ | 1819132257053442048 |
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| author | Nienke R. Wevers Dhanesh G. Kasi Taylor Gray Karlijn J. Wilschut Benjamin Smith Remko van Vught Fumitaka Shimizu Yasuteru Sano Takashi Kanda Graham Marsh Sebastiaan J. Trietsch Paul Vulto Henriëtte L. Lanz Birgit Obermeier |
| author_facet | Nienke R. Wevers Dhanesh G. Kasi Taylor Gray Karlijn J. Wilschut Benjamin Smith Remko van Vught Fumitaka Shimizu Yasuteru Sano Takashi Kanda Graham Marsh Sebastiaan J. Trietsch Paul Vulto Henriëtte L. Lanz Birgit Obermeier |
| author_sort | Nienke R. Wevers |
| collection | DOAJ |
| description | Abstract Background Receptor-mediated transcytosis is one of the major routes for drug delivery of large molecules into the brain. The aim of this study was to develop a novel model of the human blood–brain barrier (BBB) in a high-throughput microfluidic device. This model can be used to assess passage of large biopharmaceuticals, such as therapeutic antibodies, across the BBB. Methods The model comprises human cell lines of brain endothelial cells, astrocytes, and pericytes in a two-lane or three-lane microfluidic platform that harbors 96 or 40 chips, respectively, in a 384-well plate format. In each chip, a perfused vessel of brain endothelial cells was grown against an extracellular matrix gel, which was patterned by means of surface tension techniques. Astrocytes and pericytes were added on the other side of the gel to complete the BBB on-a-chip model. Barrier function of the model was studied using fluorescent barrier integrity assays. To test antibody transcytosis, the lumen of the model’s endothelial vessel was perfused with an anti-transferrin receptor antibody or with a control antibody. The levels of antibody that penetrated to the basal compartment were quantified using a mesoscale discovery assay. Results The perfused BBB on-a-chip model shows presence of adherens and tight junctions and severely limits the passage of a 20 kDa FITC-dextran dye. Penetration of the antibody targeting the human transferrin receptor (MEM-189) was markedly higher than penetration of the control antibody (apparent permeability of 2.9 × 10−5 versus 1.6 × 10−5 cm/min, respectively). Conclusions We demonstrate successful integration of a human BBB microfluidic model in a high-throughput plate-based format that can be used for drug screening purposes. This in vitro model shows sufficient barrier function to study the passage of large molecules and is sensitive to differences in antibody penetration, which could support discovery and engineering of BBB-shuttle technologies. |
| first_indexed | 2024-12-22T09:28:31Z |
| format | Article |
| id | doaj.art-9a670cc2537543efb3a82377ecf0d4a8 |
| institution | Directory Open Access Journal |
| issn | 2045-8118 |
| language | English |
| last_indexed | 2024-12-22T09:28:31Z |
| publishDate | 2018-08-01 |
| publisher | BMC |
| record_format | Article |
| series | Fluids and Barriers of the CNS |
| spelling | doaj.art-9a670cc2537543efb3a82377ecf0d4a82022-12-21T18:31:00ZengBMCFluids and Barriers of the CNS2045-81182018-08-0115111210.1186/s12987-018-0108-3A perfused human blood–brain barrier on-a-chip for high-throughput assessment of barrier function and antibody transportNienke R. Wevers0Dhanesh G. Kasi1Taylor Gray2Karlijn J. Wilschut3Benjamin Smith4Remko van Vught5Fumitaka Shimizu6Yasuteru Sano7Takashi Kanda8Graham Marsh9Sebastiaan J. Trietsch10Paul Vulto11Henriëtte L. Lanz12Birgit Obermeier13Mimetas BVMimetas BVBiogenMimetas BVBiogenMimetas BVYamaguchi University Graduate School of MedicineYamaguchi University Graduate School of MedicineYamaguchi University Graduate School of MedicineBiogenMimetas BVMimetas BVMimetas BVBiogenAbstract Background Receptor-mediated transcytosis is one of the major routes for drug delivery of large molecules into the brain. The aim of this study was to develop a novel model of the human blood–brain barrier (BBB) in a high-throughput microfluidic device. This model can be used to assess passage of large biopharmaceuticals, such as therapeutic antibodies, across the BBB. Methods The model comprises human cell lines of brain endothelial cells, astrocytes, and pericytes in a two-lane or three-lane microfluidic platform that harbors 96 or 40 chips, respectively, in a 384-well plate format. In each chip, a perfused vessel of brain endothelial cells was grown against an extracellular matrix gel, which was patterned by means of surface tension techniques. Astrocytes and pericytes were added on the other side of the gel to complete the BBB on-a-chip model. Barrier function of the model was studied using fluorescent barrier integrity assays. To test antibody transcytosis, the lumen of the model’s endothelial vessel was perfused with an anti-transferrin receptor antibody or with a control antibody. The levels of antibody that penetrated to the basal compartment were quantified using a mesoscale discovery assay. Results The perfused BBB on-a-chip model shows presence of adherens and tight junctions and severely limits the passage of a 20 kDa FITC-dextran dye. Penetration of the antibody targeting the human transferrin receptor (MEM-189) was markedly higher than penetration of the control antibody (apparent permeability of 2.9 × 10−5 versus 1.6 × 10−5 cm/min, respectively). Conclusions We demonstrate successful integration of a human BBB microfluidic model in a high-throughput plate-based format that can be used for drug screening purposes. This in vitro model shows sufficient barrier function to study the passage of large molecules and is sensitive to differences in antibody penetration, which could support discovery and engineering of BBB-shuttle technologies.http://link.springer.com/article/10.1186/s12987-018-0108-3Blood–brain barrierMicrofluidicsOrgan-on-a-chipBBBAntibody transcytosis |
| spellingShingle | Nienke R. Wevers Dhanesh G. Kasi Taylor Gray Karlijn J. Wilschut Benjamin Smith Remko van Vught Fumitaka Shimizu Yasuteru Sano Takashi Kanda Graham Marsh Sebastiaan J. Trietsch Paul Vulto Henriëtte L. Lanz Birgit Obermeier A perfused human blood–brain barrier on-a-chip for high-throughput assessment of barrier function and antibody transport Fluids and Barriers of the CNS Blood–brain barrier Microfluidics Organ-on-a-chip BBB Antibody transcytosis |
| title | A perfused human blood–brain barrier on-a-chip for high-throughput assessment of barrier function and antibody transport |
| title_full | A perfused human blood–brain barrier on-a-chip for high-throughput assessment of barrier function and antibody transport |
| title_fullStr | A perfused human blood–brain barrier on-a-chip for high-throughput assessment of barrier function and antibody transport |
| title_full_unstemmed | A perfused human blood–brain barrier on-a-chip for high-throughput assessment of barrier function and antibody transport |
| title_short | A perfused human blood–brain barrier on-a-chip for high-throughput assessment of barrier function and antibody transport |
| title_sort | perfused human blood brain barrier on a chip for high throughput assessment of barrier function and antibody transport |
| topic | Blood–brain barrier Microfluidics Organ-on-a-chip BBB Antibody transcytosis |
| url | http://link.springer.com/article/10.1186/s12987-018-0108-3 |
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