Fig. 1: Generation of Hi-Q brain organoids.

A Microscopic view showing a custom-designed spherical plate with microwells and schematic view of the inverted pyramid-like microwell. The angles and measurements are shown. B hiPSC settles and readily forms spheres in the spherical plate’s microwells. The panel shows the scale bar. C Formation of neurospheres in the microwells of the spherical plate. The magnified image at the right shows a representative neurosphere. Magnified panels show a neural rosette stained with actin (green) with primary cilia emanating into the lumen (L) at the apical side of the rosette marked by Arl13B (Red). Panels show the scale bar. D Different stages of Hi-Q brain organoid generation. E A group of neurospheres. The graph at right shows no significant difference between the organoid diameter across four independent batches. At least twenty (n = 20) randomly chosen neurospheres were measured from each batch. Statistical analysis was done using one-way ANOVA, followed by Tukey’s multiple comparisons test. Data presented as mean ± SEM. The cell line used is IMR90. The panel shows the scale bar. F Maturation of neurospheres into Hi-Q brain organoids in spinner flasks. Macroscopic images show a group of organoids. The panel shows the scale bar. G Hi-Q brain organoids increase in size progressively over time from day 15 to day 150. The panel shows the scale bar. H The average diameter of twenty-day (i) and fifty-day (ii) old organoids were differentiated from four independent hiPSC lines. Note that there is no significant difference among the different hiPSC donors within each time point. At least twenty-five (n = 25) randomly chosen organoids were analyzed across three independent batches (N = 3). Statistical analysis was done using one-way ANOVA, followed by Tukey’s multiple comparisons test. Data presented as mean ± SEM. I The graph shows no size difference between twenty-day and sixty-day-old organoids, showing a regulated growth rate between two-time points. At least twenty-five (n = 25) randomly chosen organoids were analyzed across three independent batches (N = 3). Data distribution is given in the panel (H). Statistical analysis was done using one-way ANOVA, followed by Tukey’s multiple comparisons test. Data presented as mean ± SEM. J The bar diagram quantifies the number of disintegrated organoids in each batch. At least three hundred (n = 300) organoids were randomly sampled across four independent batches (N = 4).