Fig. 6

Bioengineered vessels with astrocytes have reduced Aβ42 accumulation compared to bioengineered vessel lacking astrocytes and astrocyte-derived apoE in tissue chamber media reduces Aβ42 binding to collagen-I. a Immunofluorescent staining confirms the formation of an endothelial monolayer (CD31) surrounded by several layers of SMC (α-SM actin) with astrocytes genotyped as APOEε3/3 on the antelumen (GFAP). b 1 μM Aβ42 was injected into the tissue chamber of bioengineered vessels without (bipartite) or with astrocytes (tripartite). After 24 h, tissues were lysed in RIPA and Aβ42 accumulation was quantified using ELISA. c apoE levels in bipartite or tripartite bioengineered vessels were measured in RIPA lysate by immunoblot and normalized to GAPDH. d Astrocyte-secreted apoE levels in the circulation and chamber media were measured by ELISA. e Black 96-well plates were coated either with 50 μg/mL rat-tail collagen-I or 10% BSA for 24 h before incubating with a solution of 1 μM of FITC-Aβ42 in conditioned media from the chamber or circulating media from bipartite or tripartite bioengineered vessels. After 24 h and extensive washes, fluorescence was measured at 520 nm, excitation 490 nm. Points in graphed data represent individual bioengineered vessels, bars represent mean, error bars represent ±SEM and analysed by Student’s t-test or one way ANOVA *P < 0.05 and **P < 0.01. L: lumen