Fig. 1
Expression and localization of marker proteins in hiPSC-derived brain cell types of the NVU. HiPSCs were derived from a late-onset Alzheimer disease patient (LOAD NVU model) and a healthy elderly control subject (CON NVU model). (A) Flow cytometry confirmed high expression of cell type-specific phenotypic markers in pericytes (PCs), astrocytes (ACs), neural stem cells (NSCs; all mean ± SD; n ≥ 3), and microglia-like cells (MGCs; mean ± SD; n = 2). Representative immunofluorescence images confirmed protein expression of (B) NG2 and αSMA in PCs, scale bar 200 μm, (C) EAAT1 and EAAT2 in ACs, scale bar 100 μm, (D) CD45 and AIF1 in MGCs, scale bar 100 μm, (E) PAX6, SOX1, and NES in NSCs, scale bar 100 μm, and (F) GLUT1/SLC2A1, ZO1, OCLN, and CLDN5 in BCECs, scale bar 100 μm. (G) Measurement of transendothelial electrical resistance (TEER) and sodium fluorescein (NaF) permeability to analyze the integrity of the blood-brain barrier in CON and LOAD. TEER (left) and permeability coefficient (PCNaF) (right) values are shown for mono- and co-cultures consisting of BCECs with or without PCs, ACs, MGCs, or NSCs. Box (median and lower/upper quartile) and whisker (minimum/maximum) plots display n = 4–6 independent biological assays (no outliers), one-tailed t-test, *p < 0.05