Fig. 3.

In vivo-like choroid plexus capillaries and epithelium as a blood-CSF barrier (BCSFB) in the engineered microenvironment to reconstruct the human ChP. a Confocal images of immunostainned nuclei (blue), CD31 (red), and occludin (magenta) expressed on the microvessels cultured in each condition (scale bar: 50 μm). b RT-qPCR results of occludin expressed in 3D cell cultured HBMECs and HBVPs in each cell culture condition (-laminin, +laminin, static, and dynamic) (n = 3 for each condition, *p < 0.05, **p < 0.01, and ****p < 0.0001 by student t-test). c Microvascular density of microvessels cultured in each culture condition (n = 6 for each condition, **p < 0.01 by student t-test). d Representative confocal image of nuclei (blue), αSMA (green), CD31 (red) expressed on the in vivo-like ChP capillaries (scale bar: 100 μm). e Beads flow in the perfusable microvessels cultured in the engineered microenvironment (blood vessels, yellow; 0.2 μm microbeads, red). f Immunostaining of f-actin (green), FoxJ1 (red), RSPH9 (yellow) expressed on HCPEpiCs cultured in the ChP epithelium-on-a-chip in each culture condition. (scale bar: 50 μm) g Quantification of the coverage of the HCPEpiCs (n = 6 for each condition, *p < 0.05, **p < 0.01, and ****p < 0.0001 by student t-test). h qRT-PCR results RSPH9 expressed on the epithelium by varying ECM and flow conditions (n = 3 for each condition, *p < 0.05 and ***p < 0.001 by student t-test). i-j (i) qRT-PCR results (n = 3 for each condition, *p < 0.05, and **p < 0.01 by student t-test) and (j) confocal images of nuclei (blue) and claudin-1 (magenta) expressed on the epithelium by varying ECM and flow conditions (lam: laminin) (scale bar: 20 μm). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)