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. 2023 May 30;8:27. doi: 10.1038/s41536-023-00301-7

Fig. 2. Survival and differentiation and functions of grafted human cerebral organoids in the infarct core of stroke mice.

Fig. 2

a Design of the experiments for b–m. Representative images showing the coronal section stained with STEM123 and GFAP in animal A (b) or with NeuN and GFAP in animal B (c) on d 180 after organoids transplantation, in which, b1 and c1 showing infarct area (scale bar, 200 μm), b2 and c2 showing the survival of organoids (scale bar, 200 μm), the boxed area in b2 and c2 was shown in high magnification in b3 and c3 respectively (scale bar, 50 μm), the boxed area in b3 and c3 was shown in high magnification in b4 and c4 respectively (scale bar, 5 μm). Representative images showing HN+ graft cells co-labeled with NeuN (d) or GFAP (e) (left, scale bar, 200 μm) on d 180 after transplantation. The boxed area in leftward image was shown in high magnification on the middle image (scale bar, 50 μm), and the boxed area in middle image was shown in high magnification on the right three panels (scale bar, 5 μm). f–h Bar graphs show cerebral organoids’ cellular composition at 180 d after transplantation. n = 6 animals. More than 4,500 cells from random fields were manually counted in each condition. i Representative image showing organoids neurons (GFP+) and astrocytes (GFAP+) (upper, scale bar, 50 μm) and high magnification images from a selected area in the upper image (lower, scale bar, 20 μm) at 180 d after transplantation. j Images showing whole-cell patch-clamp recordings from organoids neurons (GFP+). Black dotted lines indicate electrode. Scale bar, 40 μm. k Voltage traces showing APs from GFP+ neuron in response to current-step, in which, red traces showing the first AP induced by 0 pA injected current. l Number of APs evoked by various current steps from GFP+ neurons. n = 13 neurons from 5 mice. m Representative traces of sPSCs from the organoids neurons. n = 34 neurons from 7 mice. In b–e, the area outlined by the yellow dotted line indicates the infarct core. In f–h and l, data were presented as mean ± standard deviation. HO Hoechst 33258 dye, HN Human Nuclei.