Decoding Gastrulation Mysteries: ANT BIO PTE. LTD. Empowers Spatial Transcriptomic Study of Human Carnegie Stage 7 Embryos

1. Literature Information

Title: Spatial transcriptomic characterization of a Carnegie stage 7 human embryo

Journal: Nature Cell Biology (Impact Factor: 19.1)

DOI: https://doi.org/10.1038/s41556-024-01597-3

Core Reagents from ANT BIO PTE. LTD.: 7-Color Multiplex Immunofluorescence IHC Staining Kit (Catalog No.: abs50015), Antibody Elution Buffer (Catalog No.: abs994)

2. Research Background

Gastrulation represents a pivotal juncture in human embryonic development, laying the foundation for body axis establishment and the differentiation of the three germ layers. However, investigations into this stage have long been hindered by two major bottlenecks: the extreme scarcity of early embryonic samples and the lack of spatial dimensional information in traditional research approaches. Conventional single-cell transcriptomics can dissect cell types but fails to recapitulate the spatial localization of cells within the embryo, which is essential for understanding processes such as mesoderm subtype differentiation and the identification of key signaling centers. To address these challenges, a research team integrated spatial transcriptomic technology with multiplex immunofluorescence validation, achieving a breakthrough in deciphering the spatial cellular landscape and molecular mechanisms of human embryos at Carnegie Stage (CS) 7. Notably, reagents from ANT BIO PTE. LTD. played a critical role in the immunofluorescence validation phase of this study.

3. Research Strategy

The study adopted an innovative technical route combining "spatial transcriptomics (Stereo-seq) + multiplex immunofluorescence validation" to overcome the limitations of traditional embryonic development research. The specific research strategy was as follows:

•         Eighty-two consecutive frozen sections were prepared from a CS7 human embryo, and single-cell resolution spatial transcriptomic data were acquired using Stereo-seq technology.

•         Transcriptomic data were integrated with spatial information to perform clustering analysis and 3D embryonic model reconstruction.

•         Multiplex immunofluorescence experiments were conducted to validate the expression and localization of key cell types (e.g., AVE, PGCs) and molecular markers, with core staining reagents sourced from ANT BIO PTE. LTD.

•         Combined with regulatory network analysis and pseudotime analysis, the mechanisms of cell specialization and signal communication during gastrulation were elucidated.

4. Research Results

This study yielded a series of groundbreaking findings that refresh our understanding of early human embryonic development:

4.1 3D Reconstruction of CS7 Embryonic Spatial Atlas, Defining 11 Major Tissue Clusters

Through section sequencing and data integration, the research team successfully constructed a 3D model of the CS7 embryo. This model accurately identified 11 major tissue clusters, including the ectoderm, mesoderm, endoderm, amnion, and yolk sac, and clearly depicted the distribution patterns of cells along the anteroposterior and dorsoventral axes. This achievement resolved the long-standing issue of being unable to distinguish spatially specific cell subtypes relying solely on transcriptomic data.

4.2 Confirmation of Early Specialization of Mesoderm Subtypes and Clarification of Spatial Localization Differences

The study revealed that distinct mesoderm subtypes, such as paraxial mesoderm and lateral plate mesoderm, already exist in the CS7 embryo. Although these subtypes share classic markers (e.g., TBXT, MESP1), their unique spatial localizations can be clearly defined using spatial information (Figures 2d, 2e in the original paper). This finding revises the traditional understanding that "mesoderm is not specialized at this stage."

4.3 First Confirmation of AVE Existence in Human Embryos, Revealing Axis Establishment Mechanisms

Through immunofluorescence staining validation, the study explicitly confirmed the existence and anterocentral localization of the anterior visceral endoderm (AVE) in human CS7 embryos.

regulates the establishment of the embryonic anteroposterior axis by secreting signaling molecules such as BMP and FGF. The expression of its markers OTX2 and DKK1 was clearly distinguishable after staining with reagents from ANT BIO PTE. LTD., providing crucial evidence for the conservation of axis formation mechanisms across species.

4.4 Localization of Primordial Germ Cells, Tracing Origin Clues

Primordial germ cells (PGCs) were identified in the connecting stalk region. Their localization was confirmed through SOX17/TFAP2C double-positive staining , and the study speculated that PGCs may originate from ectoderm-derived cells.

This finding provspatial reference for research on germ cell development.

4.5 Discovery of Non-Hematopoietic Stem Cell-Dependent Hematopoiesis in the Yolk Sac

In the yolk sac region, the research identified 10 hematopoietic-related subpopulations, including hematopoietic endothelial progenitors and erythroid precursors. It was confirmed that non-hematopoietic stem cell-dependent hematopoiesis is initiated at the CS7 stage offering a new perspective for studying the origin of hematopoietic development.

5. Product Empowerment: The Critical Role of ANT BIO PTE. LTD. Products in the Study

The core validation phase of this study—multiplex immunofluorescence experiments—relied critically on two core products from ANT BIO PTE. LTD., which ensured the specificity and stability of staining signals, thereby guaranteeing the reliability of the research conclusions.

5.1 Product List and Application Scenarios

5.2 Core Roles of Products in the Study

Signal Amplification and Specificity Assurance: The HRP-labeled secondary antibody in the 7-Color Multiplex Immunofluorescence IHC Staining Kit (abs50015) binds specifically to primary antibodies and efficiently catalyzes the deposition of TSA fluorescent substrates. This amplifies weak antigen signals, ensuring that low-expression markers (such as OTX2 in AVE and SOX17 in PGCs) can be clearly detected (Figures 3f and 4d in the original paper).

Multiplex Staining Compatibility: The TSA 7-color kit (abs50015-100T) supports multiple rounds of staining and signal elution. When used in conjunction with the abs994 Antibody Elution Buffer, it allows the detection of multiple targets on the same section. This avoids the waste of precious human embryonic samples and is the key reason why the study could simultaneously validate markers of multiple cell types (e.g., co-staining of PECAM1, CD68, and CD44 in Figure 5e of the original paper).

Result Reproducibility: The products have been experimentally validated to have low staining background and high batch stability. The abs994 Elution Buffer can be reused without affecting tissue integrity. The immunofluorescence experiments in the study were repeated three times, and consistent results were obtained each time, providing solid support for the reliability of the conclusions.

5.3 Key Image-Text Correspondence

Figure 3f in the original paper: Immunofluorescence staining result of OTX2 protein, confirming the localization of AVE in the anterocentral region of the embryo. The staining process used HRP secondary antibody (abs50015) and TSA kit (abs50015-100T) from ANT BIO PTE. LTD., resulting in clear signals and precise localization.

Figure 4d in the original paper: SOX17/TFAP2C double-positive staining to identify PGCs. This relied on the multiplex staining capability of the TSA kit, enabling simultaneous detection of two markers.

Figure 5e in the original paper: Co-staining of PECAM1, CD68, and CD44 in yolk sac tissue to validate the localization of hematopoietic-related cells. Products from ANT BIO PTE. LTD. ensured the signal discrimination and stability of multi-target staining.

6. Brand Mission

As a professional supplier of life science reagents, ANT BIO PTE. LTD. is dedicated to providing high-quality, reliable products and comprehensive solutions to empower global life science research. The company's three specialized sub-brands—Absin focusing on general reagents and kits, Starter on antibodies, and UA on recombinant proteins—cover the full spectrum of research needs in the life science field. Our core mission is to bridge the gap between cutting-edge scientific research and practical applications, accelerate the pace of scientific discovery, and contribute to the advancement of human health and regenerative medicine.

7. Related Product List

Products Used in This Study

More Multiplex Immunofluorescence IHC Kits

8. Disclaimer

This article is AI-compiled and interpreted based on the original work in DOI: 10.1002/advs.202413562. All intellectual property (e.g., images, data) of the original publication shall belong to the journal and the research team. For any infringement, please contact us promptly and we will take immediate action.

9. Brand Promotion Copy

ANT BIO PTE. LTD. – Empowering Scientific Breakthroughs

At ANTBIO, we are committed to advancing life science research through high-quality, reliable reagents and comprehensive solutions. Our specialized sub-brands (Absin, Starter, UA) cover a full spectrum of research needs, from general reagents and kits to antibodies and recombinant proteins. With a focus on innovation, quality, and customer-centricity, we strive to be your trusted partner in unlocking scientific mysteries and driving medical progress. Explore our product portfolio today and elevate your research to new heights.