High-resolution transcriptional and morphogenetic profiling of cells from micropatterned human ESC gastruloid cultures

• Published in: eLife Vol. 9
• Main Authors: Minn, Kyaw Thu, Fu, Yuheng C, He, Shenghua, Dietmann, Sabine, George, Steven C, Anastasio, Mark A, Morris, Samantha A, Solnica-Krezel, Lilianna
• Format: Journal Article
• Language: English
• Published: England eLife Sciences Publications, Ltd 18.11.2020; eLife Sciences Publications Ltd
• Subjects: amnion; Basic Helix-Loop-Helix Transcription Factors - genetics; Basic Helix-Loop-Helix Transcription Factors - metabolism; Body Patterning; Bone Morphogenetic Protein 4 - pharmacology; Cell Adhesion Molecules - genetics; Cell Adhesion Molecules - metabolism; Cell Culture Techniques; Cell Differentiation - drug effects; Cell Differentiation - physiology; Cell Line; cell sorting; Cells, Cultured; Developmental Biology; Embryonic Stem Cells - physiology; Gastrula - cytology; gastrulation; Gene Expression Regulation, Developmental; germ layers; Humans; primordial germ cells; Stem Cells and Regenerative Medicine; Transcription Factor AP-2 - genetics; Transcription Factor AP-2 - metabolism; Transcription, Genetic - physiology; trophectoderm; Wnt Proteins - genetics; Wnt Proteins - metabolism; germ layers; stem cells; trophectoderm; cell sorting; developmental biology; regenerative medicine; primordial germ cells; human; gastrulation; amnion
• ISSN: 2050-084X; 2050-084X
• DOI: 10.7554/eLife.59445

During mammalian gastrulation, germ layers arise and are shaped into the body plan while extraembryonic layers sustain the embryo. Human embryonic stem cells, cultured with BMP4 on extracellular matrix micro-discs, reproducibly differentiate into gastruloids, expressing markers of germ layers and extraembryonic cells in radial arrangement. Using single-cell RNA sequencing and cross-species comparisons with mouse, cynomolgus monkey gastrulae, and post-implantation human embryos, we reveal that gastruloids contain cells transcriptionally similar to epiblast, ectoderm, mesoderm, endoderm, primordial germ cells, trophectoderm, and amnion. Upon gastruloid dissociation, single cells reseeded onto micro-discs were motile and aggregated with the same but segregated from distinct cell types. Ectodermal cells segregated from endodermal and extraembryonic but mixed with mesodermal cells. Our work demonstrates that the gastruloid system models primate-specific features of embryogenesis, and that gastruloid cells exhibit evolutionarily conserved sorting behaviors. This work generates a resource for transcriptomes of human extraembryonic and embryonic germ layers differentiated in a stereotyped arrangement.