Fig. 2: iPSCs derived cerebral organoid formation process.

a Generation steps. b Percentage of organoid harvestability results (shaker versus other systems, *p = 0.0103 for static, **p = 0.0054 for a spinner, ns; p = 0.5873 for RCCS and ns; p = 0.4343 for µ-platform, n = 2 for independent replicates = 2). Maturation steps by c static and dynamic systems as d shaker, e spinner, f RCCS microgravity bioreactor, and g µ-platform. From left to right, the system overviews, macroscopic images, bright-field microscopic images (white arrows indicate off-target cell differentiation in preserved matrigel matrix) (Zeiss Axio Vert.A1) of organoids and H&E stained (Motic, AE31E) organoids matured in different systems at day 60 (white arrows indicate sequentially organized cell nucleus and neural rosette-like structures) (scale bars = 200 μm for bright-field and 100 μm for H&E staining images, independent replicates = 3), and plots of organoid sizes (day 120 versus day 60, ***p = 0.0002 for static, ns; p = 0.9582 for shaker, ns; p = 0.5787 for a spinner, ns; p = 0.9945 for RCCS and ns; p = 0.9918 for µ-platform, independent replicates = 2) where diameters were measured with the ImageJ program at days 0, 30, 60, and 120 (n = 40, 33, 20, 10 for static, n = 40, 40, 34, 24 for shaker, n = 40, 36, 16, 8 for a spinner, n = 40, 40, 30, 18 for RCCS, and n = 40, 38, 28, 18 for µ-platform, all for on day 0, 30, 60, 120, respectively).