Fig. 7: Hi-Q brain organoids allow glioma invasion assays and can adapt medium-throughput drug screening assays to identify anti-glioma compounds.

A Experimental scheme for a glioma invasion assay using Hi-Q brain organoids that can be adapted for drug screening. B Hi-Q brain organoids allow GSC invasion as spheres (i) or single cells (ii). The magnified panel at the right shows invading GSCs with their typical protrusions into the organoid tissues (arrows). The bar diagram at the right quantifies the depth invaded by GSCs. Shown are representative images from at least six experiments from three independent batches of organoids. Panels show the scale bar. C, D Assay design adapted to screen the compounds preventing GSC invasion. Representative images showing invading GSC into Hi-Q brain organoids (C). At least 12 organoids (n = 12) from three independent experiments were analyzed. Statistical analysis was carried out by One-way ANOVA, followed by Tukey’s multiple comparisons test, ***P < 0.001. Data presented as mean ± SEM. Automated imaging and computer-based counting of GSC spots in each organoid. Two organoids were tested for each compound. The algorithm assigns a space when there are no invading GSCs. Dark blue denotes more GSC spots in organoids. Each box represents a well that is supplied with a compound (D). E Dot plot shows the selected compounds that prevent GSC invasion into organoids. Each dot denotes the number of GSC spots in organoids. The table at the right lists selected ten compounds that can stop the GSCs’ invasions into brain organoids. The biological functions of each compound are also listed. F Quantitative 3D imaging of GSCs invasion into brain organoids. The top panel shows the GSC invasion from spheres. The bottom panel shows GSCs invasions from GSCs suspension. Fulvestrant and Selumtinib significantly prevent GSC invasion into brain organoids as selected compounds. The invaded GSC (as spheres or single cells) were marked with red arrowheads. White arrowheads point to the spheres or cells that fail to invade the organoids in the presence of drugs. Panels show the scale bar. The bar diagram quantifies the inhibitory effects of the selected compounds on GSC invasion. Average measurements from at least ten organoids (n = 10) per drug condition were considered. Statistical analysis was carried out by One-way ANOVA, followed by Tukey’s multiple comparisons test, ***P < 0.001, **P < 0.01. Data presented as mean ± SEM.