Fig. 3

AD-associated phenotypes observed in patient-derived 3D co-cultures. A Representative bright field images of HC and AD co-cultures. Scale bars, 100 μm. B Survival of HC (n = 12) and AD (n = 13) MDMi in 2D and 3D mono-cultures, and 3D co-cultures. Arrows and values indicate the fold change increase of the mean survival in 3D and 3D co-cultures versus 2D in each cohort. C Gene expression of the pro-apoptotic marker BAX in HC (n = 12) and AD (n = 13) 3D co-cultures. D Immunostaining against Iba1 (green) and Tubb3 (red) and 3D surface reconstruction of a 3D co-culture. Areas of contact between Iba1 + (MDMi) and Tubb3 + (ReNcell VM) cells are highlighted in yellow (white arrows). Scale bars, 100 μm. Quantification of E contact area and F number of contacts between Iba1 + and Tubb3 + /Gfap + cells in HC (n = 6) and AD (n = 6) 3D co-cultures. Concentration of G platelet-derived growth factors AA (PDGF-AA), H erythropoietin (EPO), I interferon-γ (IFN-γ), J angiopoietin 2 and K granulocyte–macrophage colony stimulating factor (GM-CSF) secreted by HC (n = 8–10) and AD (n = 9–10) MDMi in 3D mono-cultures and co-cultures. Baseline secretion by 3D ReNcell VM mono-cultures is represented with a dotted black line. L Representative images of 3D co-cultures containing FITC-Aβ aggregates. MDMi exhibit a resting, ramified morphology at 0 h. They progressively become polarized and acquire an activated, round morphology with enlarged soma upon reaching and clustering on the Aβ deposit at 36 h and 48 h (white arrows). Scale bars, 100 μm. See also Additional file 10: Movie S3. M Total surveillance distance, N speed and O clustering around Aβ aggregates of HC (n = 5) and AD (n = 5) MDMi in 3D co-cultures. P Concentration of pro-inflammatory cytokines IL-6, IL-1β and IL-18 secreted by untreated (UT) and Aβ-treated HC (n = 10 UT cultures; n = 7 Aβ-treated cultures) and AD (n = 10 UT cultures; n = 7 Aβ-treated cultures) MDMi in 3D co-cultures. Data are presented as mean ± SD. Data points and n represent biological replicates (donors). One-way ANOVA with Tukey’s multiple comparison test in B; unpaired Student’s t test with or without Welch’s correction, two-tailed in M, P; Mann–Whitney test, two-tailed in E, F; two-way ANOVA with Tukey’s multiple comparison test in G–K; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001