hMGEOs Recapitulate Human MGE Development(A-C) Immunostaining for SOX2 and GFP reveals the diminishment of VZ-like area in hMGEOs in a long-term culture. Scale bar, 50 μm.(D) Thickness quantification of VZ-like area in hMGEOs after 22 (n=21), 44 (n=32), and 71 (n=19) days of culture. The maximum diameter of each VZ-like area (indicated with arrows in the schematic diagram on the right) was used for quantification. Mean ± SD are shown for each stage. ***, P<0.001.(E and F) Immunostaining for GFP, SOX2, and DLX2 of day 22 (E) and day 71 (F) hMGEOs sections. Scale bar, 25 μm.(G) Immunostaining for vGAT and GABA in 80 day old organoid section. Scale bar, 25 μm.(H) Schematic illustrating the modification of hMGEO protocol to test effect of various SHH doses on dorsal-ventral patterning.(I) Quantitative RT-qPCR analysis of NKX2-1 and SST expression. Mean ± SD are shown for each condition (n=4). **, P<0.01; *, P<0.05.(J) Immunostaining for SST in 46 day old hMGEO sections. Scale bar, 25 μm.(K-M) Representative images showing migrating streaks interior of 75 day old hMGEOs. The migration directions are indicated with arrows. Immunostaining was performed for SOX2, NRP1, CXCR4, and GFP. Scale bar, 50 μm in K, 25 μm in L and M.See also Figure S1.
