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eLifeDevelopmental BiologyPeer-reviewed journal article

Modelling the impact of decidual senescence on embryo implantation in human endometrial assembloids

Thomas M. RawlingsiD, Komal MakwanaiD, Deborah M. TayloriD, Matteo A. MolèiD, Katherine J. Fishwick, Maria TryfonosiD, Joshua OdendaaliD, Amelia HawkesiD, Magdalena Zernicka‐GoetziD, Geraldine M. HartshorneiD, Jan Joris BrosensiD, Emma S. LucasiD

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Abstract

Decidual remodelling of midluteal endometrium leads to a short implantation window after which the uterine mucosa either breaks down or is transformed into a robust matrix that accommodates the placenta throughout pregnancy. To gain insights into the underlying mechanisms, we established and characterised endometrial assembloids, consisting of gland organoids and primary stromal cells. Single-cell transcriptomics revealed that decidualized assembloids closely resemble midluteal endometrium, harbouring differentiated and senescent subpopulations in both glands and stroma. We show that acute senescence in glandular epithelium drives secretion of multiple canonical implantation factors, whereas in the stroma it calibrates the emergence of anti-inflammatory decidual cells and pro-inflammatory senescent decidual cells. Pharmacological inhibition of stress responses in pre-decidual cells accelerated decidualization by inhibiting senescence and mesenchymal-epithelial transition, processes involved in endometrial breakdown and regeneration, respectively. Accelerated decidualization resulted in entrapment of co-cultured human blastocysts in a largely static decidual matrix. By contrast, the presence of senescent decidual cells created a dynamic implantation environment, enabling embryo expansion and attachment, although their persistence led to gradual disintegration of assembloids. Our findings demonstrate that senescence controls endometrial fate decisions at implantation and highlight how endometrial assembloids may accelerate the discovery of new treatments to prevent reproductive failure.

In-depth summary

Claude Sonnet 4.6 · xhigh effort · 7 Nov 2026 · from the full text of version 1 (2 Mar 2021) · CC BY 4.0

Endometrial assembloids of gland organoids and stromal cells reproduce midluteal cell states by scRNA-seq. Senescent decidual cells, tuned with dasatinib, track with whether co-cultured human blastocysts expand or stall.

Decidualization in early pregnancy cannot be studied directly, though disorders such as recurrent implantation failure and recurrent pregnancy loss are attributed to its perturbation. The authors build an assembloid model of gland organoids plus primary stromal cells to test whether acute senescence is integral to implantation.

Key results

  1. ShownSingle-cell profiling of decidualized assembloids resolved epithelial and stromal subpopulations that map to specific menstrual-cycle phases in vivo. The dominant day-4 states correspond to the midluteal implantation window.

    Eleven clusters; dominant decidualized subsets EpS4 (n=434 cells) and SS3 (n=495 cells), cross-referenced to public in vivo data GSE4888. · Cellular complexity of decidualizing assembloids…

  2. ShownBoth compartments contain senescent subpopulations with distinct secretomes. Senescent epithelial cells (EpS5) induce p16 and p21 and secrete canonical implantation factors such as DPP4, GDF15 and IGFBP3; senescent decidual cells (SS5) express a broad SASP programme.

    EpS5 n=208, SS5 n=118 cells; 56 secreted-factor genes up in senescent epithelium vs 72 in senescent stroma, with only 15 shared. · Cellular complexity of decidualizing assembloids…; Discussion

  3. ShownDasatinib during decidualization almost removed senescent decidual cells, enlarged the decidual-cell population and cut transitional cells. It blunted the pre-decidual stress response and abolished CXCL8 secretion, while raising CXCL14, IL-15 and TIMP3.

    scRNA-seq: SS5 n=7; SS4 n=882, a 9-fold increase; transitional cells down 76%; stress GO terms down in SS3 (FDR P = 3.5 × 10^-5 for 'Response to wounding'); ELISA in assembloids from 4 biopsies. · Tyrosine kinase-dependent stress responses…

  4. SuggestedCellPhoneDB predicted extensive epithelial-stromal crosstalk buffered by non-selective interactions. Pre-decidual and senescent decidual cells were predicted to depend on tyrosine-kinase-linked signalling, which motivated the dasatinib experiment.

    270 significantly enriched non-integrin receptor-ligand interactions (FDR-corrected P < 0.05); only three highly selective epithelial-stromal pairs. · Receptor-ligand interactions in decidualizing…; Tyrosine kinase-dependent stress responses…

  5. SuggestedHuman blastocysts expanded in assembloids containing senescent decidual cells but failed to expand in dasatinib-pretreated assembloids, where the decidual matrix was largely static. Untreated assembloids gradually disintegrated.

    4 blastocysts per condition; expansion in untreated, none in dasatinib-pretreated; hCG did not differ between co-cultures; attachment and OCT4/GATA6 staining seen in one embryo. · Modelling the impact of decidual subpopulations…

Methods, data and reproducibility

Assembloids were built from midluteal endometrial gland organoids and primary stromal cells in collagen hydrogel. They were profiled by Drop-seq scRNA-seq with and without dasatinib, and by CellPhoneDB interaction analysis, ELISA and time-lapse co-culture with human blastocysts.

  • Assembloids from midluteal endometrial biopsies; Drop-seq scRNA-seq of undifferentiated, decidualized and dasatinib-decidualized assembloids (number of biopsies for scRNA-seq not stated) · human · n = Cell counts per subpopulation, e.g. 198 to 882
  • Medium-requirement test (qPCR of PAEP, SPP1) and ELISA time courses in assembloids · human · n = 3 biopsies (medium test and ELISA); 4 biopsies (dasatinib ELISA)
  • Public in vivo menstrual-cycle endometrial expression data used to place cell states · human · GEO: GSE4888
  • Vitrified spare human blastocysts co-cultured in assembloids with time-lapse imaging over 72 hours · human · n = 4 per condition
Methods & tools
Drop-Seq_tools-2.3.0, Seurat v3, UMAP, CellPhoneDB, Gene Ontology Consortium database, GraphPad Prism, ImageJ, Dasatinib, ELISA (DuoSet), qPCR
Software & environment
Drop-Seq_tools-2.3.0; Seurat v3; RStudio 1.2.5042; clustering used principal components 1:15 and resolution 0.6.
Reproducibility gaps
No repository or accession for the study's own scRNA-seq data and no analysis code are given; the Drop-seq link covers only the initial processing tools. Source data tables are referred to without URLs.
Limitations (4)
  • Simple assembloids lack a surface epithelium that would create distinct pre- and post-implantation microenvironments, and lack innate immune cells. · stated by the authors
  • Persistence of senescent decidual cells causes gradual disintegration of the assembloids, and the design precluded modelling of physiological implantation. · stated by the authors
  • The embryo co-culture used only 4 blastocysts per condition, with one embryo examined for attachment; no statistical test is reported for expansion. · noted in this summary
  • Dasatinib is a broad tyrosine kinase inhibitor, so effects on decidual fate and on embryo behaviour are attributed to lost senescence without a senescence-specific manipulation. · noted in this summary

Read this if you build endometrial organoid or assembloid models, or want a single-cell map of senescent and differentiated epithelial and decidual states during the implantation window.

Citations
274
FWCI
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top 1%

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