High-resolution transcriptional and morphogenetic profiling of cells from micropatterned human ESC gastruloid cultures
Kyaw Thu Minn,
Yuheng C Fu,
Shenghua He,
Sabine Dietmann,
Steven C George,
Mark A Anastasio,
Samantha A Morris,
Lilianna Solnica-Krezel
Affiliations
Kyaw Thu Minn
Department of Biomedical Engineering, Washington University, St. Louis, United States; Department of Developmental Biology, Washington University School of Medicine, St. Louis, United States
Department of Developmental Biology, Washington University School of Medicine, St. Louis, United States; Department of Genetics, Washington University School of Medicine, St. Louis, United States; Center of Regenerative Medicine, Washington University School of Medicine, St. Louis, United States
Shenghua He
Department of Computer Science & Engineering, Washington University, St. Louis, United States
Sabine Dietmann
Department of Developmental Biology, Washington University School of Medicine, St. Louis, United States; Division of Nephrology, Washington University School of Medicine, St. Louis, United States; Institute for Informatics, Washington University School of Medicine, St. Louis, United States
Steven C George
Department of Biomedical Engineering, University of California, Davis, Davis, United States
Mark A Anastasio
Department of Biomedical Engineering, Washington University, St. Louis, United States; Department of Bioengineering, University of Illinois, Urbana-Champaign, United States
Department of Developmental Biology, Washington University School of Medicine, St. Louis, United States; Department of Genetics, Washington University School of Medicine, St. Louis, United States; Center of Regenerative Medicine, Washington University School of Medicine, St. Louis, United States
Department of Developmental Biology, Washington University School of Medicine, St. Louis, United States; Center of Regenerative Medicine, Washington University School of Medicine, St. Louis, United States
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.