In vitro culture of cynomolgus monkey embryos beyond early gastrulation

Ma, Huaixiao; Zhai, Jinglei; Wang, Haifeng; Jiang, Xiangxiang; Wang, Xiaoxiao; Wang, Lin; Xiang, Yunlong; He, Xiaorong; Zhao, Zhen-Ao; Zhao, Bo; Zheng, Ping; Li, Lei; Wang, Hongmei

doi:10.1126/science.aax7890

Cited by 169 publications

(235 citation statements)

References 41 publications

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“…Our scRNA-seq data, encompassing 2,475 cells from two independent experiments with multiple gastruloids, uncovered expression of 25,271 transcripts. Comparisons of transcriptomic signatures with mouse (Pijuan-Sala et al, 2019) and cynomolgus monkey (Ma et al, 2019) gastrulae suggest that the hESC gastruloids resemble E7.25 mouse and 14 dpf cynomolgus monkey gastrulae ( Figures 3E-H), and therefore may represent early-to-mid stages of human gastrulation.…”

Section: Discussionmentioning

confidence: 90%

“…Interestingly, the ExE-like cluster also expressed amnion marker genes TFAP2A, HAND1, WNT6, and ISL1 (Ma et al, 2019;Zheng et al, 2019) ( Figure 2C). Accordingly, comparison with published datasets suggested that cells in ExE-like cluster resemble both TE and amnion cells.…”

Section: Exe-like Cluster Contains Cells With Gene Expression Profilementioning

confidence: 99%

“…Since NODAL signaling is known to posteriorize epiblast and is an evolutionarily conserved mesendoderm inducer from fish to mammals (Brennan et al, 2001;Schier, 2009), high NODAL activity is consistent with this cluster resembling posterior epiblast. By projecting mouse (Pijuan-Sala et al, 2019) or monkey (Ma et al, 2019) cell type labels onto the gastruloid data (Stuart et al, 2019), we found that the majority of cells in the EPI-like cluster match with mouse epiblast (>80%) ( Figure 3I) and monkey post-implantation epiblast (>95%) ( Figure 3J). These results suggest that the EPI-like cluster contains cells similar in transcriptomic signature to both monkey and mouse epiblast, and thus precursors of the germ layers.…”

Section: Epi-like Cluster With Characteristics Of Post-implantation Ementioning

confidence: 99%

“…Endoderm-1 expressed both primitive endoderm marker GATA6 and definitive endoderm marker FOXA2 ( Figure 2C), consistent with findings from protein expression analysis ( Figure 1D). Projection of monkey (Ma et al, 2019) cell type labels onto the gastruloid data suggested that Endoderm-1 matches with a mix of visceral endoderm (~20%), ExE mesenchyme (~15%), and early/late gastrulating cells (~20%) ( Figure 3J). Similar analysis with mouse showed Endoderm-1 matching with a mix of definitive endoderm (~20%), anterior PS (~20%), and ExE endoderm (~10%) ( Figure 3I).…”

Section: Gastruloids Contain Cells Resembling Primitive and Definitivmentioning

confidence: 99%

“…Cells in Endoderm-2 cluster were enriched for PGC markers NANOS3 and TFAP2C (Sasaki et al, 2016;Ma et al, 2019;Niu et al, 2019;Zheng et al, 2019) (Figure S2E), suggesting the presence of hPGCLCs in gastruloids. Additionally, this cluster has a similar gene expression profile to that reported for cynomolgus monkey PGCs (Sasaki et al, 2016), expressing SOX17, TFAP2C, PRDM1, POU5F1, NANOG, and T, with low or absent SOX2 level ( Figure S2G-I).…”

Section: Gastruloids Contain Cells Similar In Gene Expression Profilementioning

confidence: 99%

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High-resolution transcriptional and morphogenetic profiling of cells from micropatterned human embryonic stem cell gastruloid cultures

Fu²,

He³

et al. 2020

Preprint

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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 with radial organization. Single-cell RNA sequencing and cross-species comparisons with mouse and cynomolgus monkey gastrulae suggest that gastruloids contain seven major cell types, including epiblast, ectoderm, mesoderm, endoderm, and extraembryonic trophoblast-and amnion-like cells. 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 cells but mixed with mesodermal cells. Our work demonstrates that the gastruloid system supports 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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Section: Discussionmentioning

confidence: 90%

Section: Exe-like Cluster Contains Cells With Gene Expression Profilementioning

confidence: 99%

Section: Epi-like Cluster With Characteristics Of Post-implantation Ementioning

confidence: 99%

Section: Gastruloids Contain Cells Resembling Primitive and Definitivmentioning

confidence: 99%

Section: Gastruloids Contain Cells Similar In Gene Expression Profilementioning

confidence: 99%

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High-resolution transcriptional and morphogenetic profiling of cells from micropatterned human embryonic stem cell gastruloid cultures

Fu²,

He³

et al. 2020

Preprint

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Understanding Human Forebrain Morphogenesis and Early Expansion Using Organoids

Sutcliffe,

Lancaster

2023

Neocortical Neurogenesis in Development and Evolution

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Single‐Cell Sequencing to Unveil the Mystery of Embryonic Development

Lin

Zhong

et al. 2021

Advanced Biology

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In this process, tremendous incidences on the molecular and cellular levels take place, driving essential developmental activities such as lineage specifications, axis patterning, and organogenesis. Detailed understandings of the molecular mechanisms of this process, such as the transcriptome and epigenome, are critical for fundamental embryology study, management of reproduction-related diseases, and regenerative medicine. [2] Given that embryonic cells are scarce yet highly heterogeneous, analysis with the single-cell resolution is thus essential for a complete knowledge of embryonic development. However, conventional cell analysis, such as DNA microarray, quantitative realtime polymerase chain reaction (PCR), and sequencing, generally requires a sample of pooled cells and thus gauges the ensemble average, masking the cell heterogeneity among the tested sample. By performing analysis on individual cells separately, one could obtain analysis with single-cell resolution.The single-cell analysis normally includes four steps: 1) single-cell isolation and manipulation, 2) sample preparation, 3) sequencing, and 4) data analysis. [3] In principle, single-cell analysis can be performed following the same testing mechanisms as the bulk assays. However, there are a few challenges Embryonic development is a fundamental physiological process that can provide tremendous insights into stem cell biology and regenerative medicine. In this process, cell fate decision is highly heterogeneous and dynamic, and investigations at the single-cell level can greatly facilitate the understanding of the molecular roadmap of embryonic development. Rapid advances in the technology of single-cell sequencing offer a perfectly useful tool to fulfill this purpose. Despite its great promise, single-cell sequencing is highly interdisciplinary, and successful applications in specific biological contexts require a general understanding of its diversity as well as the advantage versus limitations for each of its variants. Here, the technological principles of single-cell sequencing are consolidated and its applications in the study of embryonic development are summarized. First, the technology basics are presented and the available tools for each step including cell isolation, library construction, sequencing, and data analysis are discussed. Then, the works that employed single-cell sequencing are reviewed to investigate the specific processes of embryonic development, including preimplantation, peri-implantation, gastrulation, and organogenesis. Further, insights are provided on existing challenges and future research directions.

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In vitro culture of cynomolgus monkey embryos beyond early gastrulation

Cited by 169 publications

References 41 publications

High-resolution transcriptional and morphogenetic profiling of cells from micropatterned human embryonic stem cell gastruloid cultures

High-resolution transcriptional and morphogenetic profiling of cells from micropatterned human embryonic stem cell gastruloid cultures

Understanding Human Forebrain Morphogenesis and Early Expansion Using Organoids

Single‐Cell Sequencing to Unveil the Mystery of Embryonic Development

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