Organization of Endothelial Cells, Pericytes, and Astrocytes into a 3D Microfluidic in Vitro Model of the Blood-Brain Barrier
| Autor: | Jack D. Wang, Mohamed E. H. ElSayed, Shuichi Takayama, Khalil Khanafer, El-Sayed Khafagy |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Cell Membrane Permeability Microfluidics Cell Culture Techniques Pharmaceutical Science Vascular permeability 02 engineering and technology In Vitro Techniques Blood–brain barrier Capillary Permeability 03 medical and health sciences Mice In vivo Drug Discovery medicine Animals ATP Binding Cassette Transporter Subfamily B Member 1 P-glycoprotein biology Tight junction Brain 021001 nanoscience & nanotechnology In vitro Coculture Techniques Cell biology 030104 developmental biology medicine.anatomical_structure Cell culture Permeability (electromagnetism) Blood-Brain Barrier Astrocytes biology.protein Molecular Medicine Endothelium Vascular 0210 nano-technology Pericytes |
| Zdroj: | Molecular pharmaceutics. 13(3) |
| ISSN: | 1543-8392 |
| Popis: | The endothelial cells lining the capillaries supplying the brain with oxygen and nutrients form a formidable barrier known as the blood-brain barrier (BBB), which exhibits selective permeability to small drug molecules and virtually impermeable to macromolecular therapeutics. Current in vitro BBB models fail to replicate this restrictive behavior due to poor integration of the endothelial cells with supporting cells (pericytes and astrocytes) following the correct anatomical organization observed in vivo. We report the coculture of mouse brain microvascular endothelial cells (b.End3), pericytes, with/without C8-D1A astrocytes in layered microfluidic channels forming three-dimensional (3D) bi- and triculture models of the BBB. The live/dead assay indicated high viability of all cultured cells up to 21 days. Trans-endothelial electrical resistance (TEER) values confirmed the formation of intact monolayers after 3 days in culture and showed statistically higher values for the triculture model compared to the single and biculture models. Screening the permeability of [(14)C]-mannitol and [(14)C]-urea showed the ability of bi- and triculture models to discriminate between different markers based on their size. Further, permeability of [(14)C]-mannitol across the triculture model after 18 days in culture matched its reported permeability across the BBB in vivo. Mathematical calculations also showed that the radius of the tight junctions pores (R) in the triculture model is similar to the reported diameter of the BBB in vivo. Finally, both the bi- and triculture models exhibited functional expression of the P-glycoprotein efflux pump, which increased with the increase in the number of days in culture. These results collectively indicate that the triculture model is a robust in vitro model of the BBB. |
| Databáze: | OpenAIRE |
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