TY - JOUR
T1 - High-resolution transcriptional and morphogenetic profiling of cells from micropatterned human esc gastruloid cultures
AU - Minn, Kyaw Thu
AU - Fu, Yuheng C.
AU - He, Shenghua
AU - Dietmann, Sabine
AU - George, Steven C.
AU - Anastasio, Mark A.
AU - Morris, Samantha A.
AU - Solnica-Krezel, Lilianna
N1 - We appreciate the technical advice on the micropattern culture from Ali Brivanlou, Eric Siggia, and their lab members. We thank Sandra Lam (Washington University School of Medicine in St. Louis) for her guidance and assistance in fabricating PDMS stamps. We thank Sarah Waye and Chuner Guo (Washington University School of Medicine in St. Louis) for constructing parts of the data curation libraries. We would also like to thank Margot Williams (Baylor College of Medicine), Mariana Beltch-eva, Gina Castelvecchi, Angela Bowman, Laura Fischer, and Thorold Theunissen (Washington University School of Medicine in St. Louis) for helpful comments on the manuscript. This study was funded by Washington University School of Medicine in St. Louis and a grant from the Children’s Discovery Institute to LSK, and a Vallee Scholar Award, an Allen Distinguished Investigator Award (through the Paul G Allen Frontiers Group), and a Sloan Research Fellowship to SAM.
We appreciate the technical advice on the micropattern culture from Ali Brivanlou, Eric Siggia, and their lab members. We thank Sandra Lam (Washington University School of Medicine in St. Louis) for her guidance and assistance in fabricating PDMS stamps. We thank Sarah Waye and Chuner Guo (Washington University School of Medicine in St. Louis) for constructing parts of the data curation libraries. We would also like to thank Margot Williams (Baylor College of Medicine), Mariana Beltch-eva, Gina Castelvecchi, Angela Bowman, Laura Fischer, and Thorold Theunissen (Washington University School of Medicine in St. Louis) for helpful comments on the manuscript. This study was funded by Washington University School of Medicine in St. Louis and a grant from the Children?s Discovery Institute to LSK, and a Vallee Scholar Award, an Allen Distinguished Investigator Award (through the Paul G Allen Frontiers Group), and a Sloan Research Fellowship to SAM. Children?s Discovery Institute Lilianna Solnica-Krezel Washington University School of Medicine in St. Louis Discretionary funds Lilianna Solnica-Krezel Vallee Foundation Vallee Scholar Award Samantha A Morris Paul G. Allen Frontiers Group Allen Distinguished Investigator Award Samantha A Morris Alfred P. Sloan Foundation Sloan Research Fellowship Samantha A Morris The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
PY - 2020/10
Y1 - 2020/10
N2 - 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.
AB - 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.
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U2 - 10.7554/eLife.59445
DO - 10.7554/eLife.59445
M3 - Article
C2 - 33206048
AN - SCOPUS:85097344925
SN - 2050-084X
VL - 9
SP - 1
EP - 34
JO - eLife
JF - eLife
M1 - e59445
ER -