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. 2022 Jul 29;13:4403. doi: 10.1038/s41467-022-32115-4

Fig. 2. Human brain organoids form a scaffolding capable of supporting neuronal microcircuitry.

Fig. 2

a Top, high-resolution, whole-section of an 8-month organoid immunostained with anti-GFAP (green), and counterstained with Hoechst (cell nuclei, blue), scale bar 1 mm. Middle, a high magnification of (top box) showing stellate appearance characteristic of astrocytes. Bottom left, Anti-GFAP-positive astrocytes (green) in an 8-month organoid co-labeled with anti-connexin 43 demonstrating gap junctions (red). Bottom right, connexin 43 gap junction (red; arrows). Middle and bottom scale bars are 40 µm. b Top, long neuronal processes labeled with anti-SMI312 (green) in an 8-month organoid, scale bar 500 µm. Bottom, anti-SMI312 (green) axons neighboring MAP2-positive (red) neurons near the margin of 8-month organoid, scale bar 20 µm. c Top, anti-GAD65 positive neurons (green) co-labeled with anti-MAP2. Bottom, Anti-Parvalbumin-positive neurons (green) in an 8-month organoid co-labeled with anti-MAP2 (red). Scale bars 40 µm. d The presynaptic marker synaptobrevin label MAP2-positive processes as puncta, scale bar 20 µm. Data from a–d were repeated independently on n = 3 organoids. e Single-cell RNA sequencing (drop-seq) shows the presence of glutamatergic neurons, GABAergic neurons and astrocyte populations. Single-cell transcriptomes (5680 cells collected from three 7-month-old organoids) are visualized as a Uniform Manifold Approximation and Projection (UMAP). Source data are provided as a Source Data File.