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. 2021 Aug 6;12:4744. doi: 10.1038/s41467-021-24776-4

Fig. 4. Organoid cultures differentiate into mesodermal lineage cells that include IBA1-expressing microglia, vasculature, and skeletal muscle.

Fig. 4

a Microglia in sensorimotor organoids immunolabeled with IBA1 at 4 weeks. Scale bar, 100 µm. b High magnification image of an IBA1+ microglial cell and TUJ1+ axon at 4 weeks of culture. Scale bar, 50 µm. c Quantification of IBA1 + cells in organoid cultures at 6 weeks. Bars indicate mean and SEM (n = 6–8 organoid cultures obtained from two independent biological differentiation replicates of each of five lines; Kruskal–Wallis, P = 0.613). d Microvasculature stained with IB4 nearby TUJ1+ neurons at 4 weeks in culture. Scale bar, 100 µm. e IB4+ microvasculature adjacent to IBA1+ microglia at 4 weeks in culture. Scale bar, 50 µm. f Electron micrograph of endothelial-lined microvessels at high (left) and low (right) magnification after 9 weeks in culture. Scale bars, 10 µm. g Elongated and striated SAA+ myotubes containing peripheral nuclei at 4 weeks. Scale bar, 100 µm. h Electron micrographs of skeletal muscle at 9 weeks showing sarcomeric organization with distinct Z lines, A and I bands, and M bands within the H zone. Scale bar, 500 nm. i Quantification of the percent of SAA+ muscle area in organoid cultures at 6 weeks. Bars indicate mean and SEM (n = 5–6 organoid cultures obtained from two independent biological differentiation replicates of each of five lines; Kruskal–Wallis, P = 0.642). Control lines: 11a, FA0000011. ALS lines: MGH5b, 19f, and FA0000012.