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Fig. 1 | BMC Biology

Fig. 1

From: Self-organising human gonads generated by a Matrigel-based gradient system

Fig. 1

The 3-LGS generates compartmentalised human gonadal organoids. A Schematic illustrating culture preparation and the 3-LGS setup. B Organoid formation in culture following testicular tissue dissociation. Images illustrate the re-organisation of gonad tissue from small cell aggregates and single cells from the same 8.5–9 wpc embryonic tissue sample. Scale bars, 500 μm. C Representative images from 8.5 wpc female and 9 wpc male embryonic gonads (PAS, periodic acid-Schiff). Female gonad accompanied by mesonephric tissue observed as small tubular lumens lined by a simple cuboidal epithelium (grey arrow, F 8.5 wpc—mesonephros inset) and larger ducts with a pseudostratified columnar epithelium (white arrow, F 8.5 wpc—mesonephros inset). Germ cells (asterisks) situated throughout the ovary, whereas localised within seminiferous cords (as indicated by white arrow, M 9 wpc zoom) in the testis. Scale bars, 50 μm. D Dissociated testicular cells reorganise into testicular organoids (TO) with distinct seminiferous-like cords (white arrows) situated within an interstitial environment after seven days in the 3-LGS and maintain structure until the end of the 14-day culture period (representative organoid images from the same 8.5–9 wpc embryonic tissue sample) (PAS, periodic acid-Schiff). E Tissue reorganisation fails to occur when mesonephros is included in the testicular tissue digest (testicular mesonephric organoid (TMO); representative organoid image from a 7.5 wpc embryonic tissue sample). The inclusion of mesonephric tissue resulted in the formation of mesonephric-like tubules throughout the interstitial space, observed as small tubular lumens lined by a simple cuboidal epithelium (grey arrow) or larger ducts with a pseudostratified columnar epithelium (white arrow; inset). The 3-LGS can also be used to generate ovarian organoids (OO) from dissociated ovarian tissue (representative organoid image from a 10 wpc embryonic tissue sample)

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