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. Author manuscript; available in PMC: 2023 Aug 1.
Published in final edited form as: Dev Biol. 2022 May 7;488:35–46. doi: 10.1016/j.ydbio.2022.05.002

Fig. 2. Making Gastruloids.

Fig. 2.

(A, B) Schematic protocol for generating mouse and human gastruloids. Pluripotent stem cells are dissociated to a single cell suspension, and an particular cell number (typically 300–500 cells) are added to each well of a 96-well plate with rounded bottoms (A). The cells will settle and aggregate to form a 3D spherical structure that, over time, begins to break symmetry, polarize its gene expression, and undergo axial elongation. The timing of exposure to agonists of Wnt signalling differs for mouse and human ES cells (B). (C) Representative microscopy images of a developing mouse gastruloid timeline between about 72 and 126 h after aggregation. Images from a light sheet movie of a gastruloid bearing ubiquitously expressed GPI linked GFP (U.Fiuza unpublished). Scale bar: 100 μm. (D, E) Sample ollections of 120m hrs old gastruloids from E14 ES (D) and P19 embryo carcinoma (E) cells.