A perfused human blood–brain barrier on-a-chip for high-throughput assessment of barrier function and antibody transport

Abstract

BackgroundReceptor-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.MethodsThe 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.ResultsThe 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).ConclusionsWe 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.Electronic supplementary materialThe online version of this article (10.1186/s12987-018-0108-3) contains supplementary material, which is available to authorized users.

Hypoxia-enhanced Blood-Brain Barrier Chip recapitulates human barrier function, drug penetration, and antibody shuttling properties

“…We also did not observe any significant difference in transcytosis ability between these two TfR antibodies when we flowed the antibodies through BBB Chips lined by primary (as opposed to iPSC-derived) human BMVECs ( Supplementary Fig. S11), which produced only a minimal (~1.4-fold) difference in transcytosis abilities for the two TfR antibodies much as previously observed in past microfluidic primary BBB models 72 . In contrast, when these anti-TfR antibodies were flowed through the vascular channel of the hypoxia-enhanced BBB Chip, we were able to demonstrate the 3-fold higher penetrance of MEM-75 into the CNS channel relative to the 13E4 antibodies that was previously predicted in vitro 70 (Fig.…”

Organ-on-a-chip: recent breakthroughs and future prospects

“…OOAC technology has developed rapidly in recent years and has enhanced our knowledge of all the major organs. Others not discussed in this review include blood vessels [99,114,115], the skin [116,117], the BBB [118,119], skeletal muscle [120,121], and the CNS [122,123].…”

Lab-On-A-Chip for the Development of Pro-/Anti-Angiogenic Nanomedicines to Treat Brain Diseases

“…In addition to the above advantages, LOCs also allow the testing of very low drug concentrations in a dose-dependent manner, even in the range of a few nanomoles per liter, which is corresponds to the therapeutic concentration [45]. Further, simultaneous, high throughput screening of different concentration on the same microfluidic platforms is possible [46,49,58]. The possibility of developing precise brain-like vasculature to perform challenging experiments has encouraged a number of researchers to adapt LOCs for their studies, Table 2.…”

“…The human-like perfusability, dynamic microenvironment, and the ability to carry out robust, rapid and reproducible assays in a controlled operational condition, with high throughput screening readouts, have made the above devices a super-tool to screen angiogenic drugs [41]. Some of the above devices were already evaluated for drug screening [32,34,36,38,40], and a few studied the effect of NPs [35,41] and NMs [39].Microfluidic technology has also been used to model brain vasculogenesis/angiogenesis [42][43][44][45][46], BBB [2,42,[47][48][49][50][51][52][53], brain tissues [54,55], and brain angiogenesis-related cellular events such as inflammation [47,50], cell migration [56], cell-cell interactions [57], etc., see Table 2. In addition, specific conditions, involving pathological-angiogenesis, such as brain tumors [46,[54][55][56]58], ischemic strokes [59], and neurodegenerative disorders including Alzheimer's disease (AD) [60], Parkinson's disease (PD) [61], and Huntington's disease (HD) [62], could also be created on-chip.In addition to this, LOCs have been developed for studying the biocompatibility, cellular uptake and transport of NMs [2,27], many of them focusing on brain angiogenesis [2,45,…”

“…Microfluidic technology has also been used to model brain vasculogenesis/angiogenesis [42][43][44][45][46], BBB [2,42,[47][48][49][50][51][52][53], brain tissues [54,55], and brain angiogenesis-related cellular events such as inflammation [47,50], cell migration [56], cell-cell interactions [57], etc., see Table 2. In addition, specific conditions, involving pathological-angiogenesis, such as brain tumors [46,[54][55][56]58], ischemic strokes [59], and neurodegenerative disorders including Alzheimer's disease (AD) [60], Parkinson's disease (PD) [61], and Huntington's disease (HD) [62], could also be created on-chip.…”