Fig. 4: Verification of the contribution of astrocyte priming to enhanced microglial Aβ42 clearance using astrocyte-microglia co-culture and glia-embedded AD cerebral organoids.
From: Astrocyte priming enhances microglial Aβ clearance and is compromised by APOE4
A Schematic overview of the experimental process for the astrocyte-microglia co-culture system. B, C After applying the priming protocol, increased Aβ42 clearance and enhanced Iba1 signal are observed in APOE3 microglia when co-cultured with APOE3 astrocytes, but not with APOE4 astrocytes. Scale bar=5 μm. D Schematic overview of the experimental process for APPswe organoids embedded with APOE3 astrocytes. E Organoids embedded with APOE3 astrocytes contain only GFAP-positive cells, with no Iba1 signal. Similar glial cell embedding was confirmed across all organoids used for the analysis. Scale bar=50 μm. F Priming does not reduce accumulated Aβ42 in organoids embedded solely with astrocytes. N = 12, from four different batches. Scale bar=50 μm. G Schematic overview of the experimental process for APPswe organoids embedded with APOE3 microglia. H Organoids embedded with APOE3 microglia contain only Iba1-positive cells, with no GFAP signal. Similar glial cell embedding was confirmed across all organoids used for the analysis. Scale bar=50 μm. I Priming does not reduce accumulated Aβ42 in organoids embedded solely with microglia. N = 12, from four different batches. Scale bar = 50 μm. J Schematic overview of the experimental process for APPswe organoids embedded with both APOE3 astrocytes and microglia. K Organoids embedded with both cell types contain GFAP-positive and Iba1-positive cells. Similar glial cell embedding was confirmed across all organoids used for the analysis. Scale bar=50 μm. L Priming reduces accumulated Aβ42 in organoids embedded with both astrocytes and microglia. N = 12, from four different batches. Scale bar=50 μm. All schematics were created in BioRender. Seo, J. (2025) https://BioRender.com/09a0byv. *p < 0.05, ***p < 0.001; ns not significant. (Two-tailed Student’s t-test). Exact P values are provided in the Source Data. Error bar ± S.E.M. test.
