Transcriptomic analysis of the interaction of choriocarcinoma spheroids with receptive vs. non-receptive endometrial epithelium cell lines: an in vitro model for human implantation
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Several in vitro systems have been reported to model human implantation; however, the molecular dynamics of the trophoblast vs. the epithelial substrate during attachment have not been described. We have established an in vitro model which allowed us to dissect the transcriptional responses of the trophoblast and the receptive vs. non-receptive epithelium after co-culture.
We established an in vitro system based on co-culture of (a) immortalized cells representing receptive (Ishikawa) or non-receptive (HEC-1-A) endometrial epithelium with (b) spheroids of a trophoblastic cell line (JEG-3) modified to express green fluorescent protein (GFP). After 48 h of co-culture, GFP+ (trophoblast cells) and GFP− cell fractions (receptive or non-receptive epithelial cells) were isolated by fluorescence-activated flow cytometry (FACS) and subjected to RNA-seq profiling and gene set enrichment analysis (GSEA).
Compared to HEC-1-A, the trophoblast challenge to Ishikawa cells differentially regulated the expression of 495 genes, which mainly involved cell adhesion and extracellular matrix (ECM) molecules. GSEA revealed enrichment of pathways related to cell division, cell cycle regulation, and metabolism in the Ishikawa substrate. Comparing the gene expression profile of trophoblast spheroids revealed that 1877 and 323 genes were upregulated or downregulated when co-cultured on Ishikawa substrates (compared to HEC-1-A), respectively. Pathways favorable to development, including tissue remodeling, organogenesis, and angiogenesis, were enhanced in the trophoblast compartment after co-culture of spheroids with receptive epithelium. By contrast, the co-culture with less receptive epithelium enriched pathways mainly related to trophoblast cell proliferation and cell cycle regulation.
Endometrial receptivity requires a transcriptional signature that determines the trophoblast response and drives attachment.
KeywordsImplantation Attachment Endometrial receptivity Transcriptomics
The authors wish to thank all members of the Basic Laboratory from Clínica EUGIN, especially Montserrat Barragán and Anna Ferrer, for critical discussion; José Buratini from Sao Paulo State University (Brasil) for critical revision of the manuscript; Camille Stephan Otto from the Biostatistics/Bioinformatics facility of the Institute for Research in Biomedicine (Barcelona) for bioinformatics analysis; Charles Pineau, Natalie Melaine, and Emmanuelle Com from Proteomics Core Facility Biogenouest (Rennes) for assistance with data analysis; and Prof. Daniel Grinberg from Universitat de Barcelona for technical support.
Paula Vergaro: experimental execution, study design, data analysis, and manuscript preparation. Gustavo Tiscornia: study design and supervision, data analysis, manuscript edition, and expert knowledge. Amelia Rodríguez: study supervision. Josep Santaló: study supervision, expert knowledge, and manuscript edition. Rita Vassena: study design and supervision, expert knowledge, and manuscript edition.
This work was supported by intramural funding of Clínica EUGIN and by the Secretary for Universities and Research of the Ministry of Economy and Knowledge of the Government of Catalonia (GENCAT 2015 DI 050).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
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