Method Profile
A human-cell BBB organoid assay estimates drug concentrations within single organoids from UPLC-MS/MS measurements of their lysates and compares uptake ratios with published in vivo brain-to-plasma data.
Explore whether quantitative uptake in a human-cell barrier model can help rank drugs for brain exposure in early development.
Three-dimensional assemblies of hCMEC/D3 human brain microvascular endothelial cells, primary pericytes and primary astrocytes; main analysis on day 3 after assembly.
Expose organoids to drugs at defined concentrations and times; quantify drug in individual organoid lysates by UPLC-MS/MS, then estimate the internal concentration using an approximate spherical organoid volume (radius 100 μm). The uptake ratio divides this estimate by the corresponding incubation concentration.
Human-cell BBB organoids
hCMEC/D3 endothelial cell line with primary pericytes and astrocytes.
UPLC-MS/MS drug assays
Drug-specific calibration and quality-control samples in organoid matrix.
Thirteen drugs were measured. Conditions entered downstream analysis when at least seven individual organoids from two preparations were quantifiable; uptake experiments were repeated independently three times. For correlation, mean uptake ratios based on at least 10 organoids were compared with published in vivo ratios after log transformation. Octreotide at background level was excluded from the logarithmic regression.
Compare measurable drug uptake ratios with heterogeneous published mammalian, primate and human brain-to-plasma data.
Thirteen drugs tested; 12 had measurable concentrations usable for the main correlation. Available drug subsets differ in primate and human analyses.
Literature brain-to-plasma or related unbound brain/CSF-to-plasma ratios, not a prospective patient cohort.
Combined mammalian Deming regression pseudo-R² 0.64; primate and human subsets 0.81 and 0.82. Four in silico comparators showed no correlation with this reference set in this study.
A Research Square preprint quantified uptake of 13 drugs in human-cell blood–brain barrier organoids. For measurable drugs, organoid uptake ratios correlated with published mammalian brain-to-plasma data, although the reference data and organoids themselves varied.
