title: Co-culture based blood-brain barrier in vitro model, a tissue engineering approach using immortalized cell lines for drug transport study.
abstract: This study evaluated the feasibility of using commercially available immortalized cell lines in building an in vitro blood-brain barrier (BBB) co-culture model for preliminary drug development studies. Astrocytes-derived acellular extracellular matrix (aECM) was introduced in the co-culture model to provide a novel biomimetic basement membrane for the endothelial cells to form tight junctions. Trans-Endothelial Electrical Resistance (TEER) and solute mass transport studies quantitatively evaluated the tight junction formation. Immuno-fluorescence microscopy and Western blot analysis qualitatively verified the expression of occludin, one of the tight junction proteins on the samples. Experimental data from a total of 13 experiments conclusively showed that the novel BBB in vitro co-culture model with aECM (CO + aECM) is promising in terms of establishing tight junction formation represented by TEER values, transport profiles, and tight junction protein expression when compared with traditional co-culture (CO) model setup or the endothelial cells cultured alone (EC). In vitro colorimetric sulforhodamine B (SRB) assay also revealed that the "CO + aECM" samples resulted in less cell loss on the basal sides of the insert membranes than traditional co-culture models. Our novel approach using immortalized cell lines with the addition of aECM was proven to be a feasible and repeatable alternative to the traditional BBB in vitro modeling.
authors.affiliation: Department of Biomedical Engineering, Florida International University, Miami, 33174, USA.
authors.forename: Zhiqi
authors.initials: Z
authors.lastname: Zhang
authors.forename: Anthony J
authors.initials: AJ
authors.lastname: McGoron
authors.forename: Eric T
authors.initials: ET
authors.lastname: Crumpler
authors.forename: Chen-Zhong
authors.initials: CZ
authors.lastname: Li
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000737:chemistry; D001692:Biological Transport; D001812:Blood-Brain Barrier / Q000378:metabolism; D015153:Blotting, Western; D002110:Caffeine / Q000378:metabolism; D002460:Cell Line; D018920:Coculture Techniques / Q000379:methods; D017097:Electric Impedance; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D005109:Extracellular Matrix / Q000378:metabolism; D005452:Fluoresceins / Q000378:metabolism; D015870:Gene Expression; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008954:Models, Biological; D062793:Occludin; D051381:Rats; D015203:Reproducibility of Results; D019108:Tight Junctions / Q000378:metabolism*; D023822:Tissue Engineering / Q000379:methods
chemical_list: D005452:Fluoresceins; D008565:Membrane Proteins; D062793:Occludin; C568986:Ocln protein, mouse; C568987:Ocln protein, rat; D002110:Caffeine; C007740:fluorexon