Rapid generation of functional nanovesicles from human trophectodermal cells for embryo attachment and outgrowth

Extracellular vesicles (EVs) are important mediators of embryo attachment and outgrowth critical for successful implantation. While EVs have garnered immense interest in their therapeutic potential in assisted reproductive technology by improving implantation success, their large-scale generation remains a major challenge. Researchers at La Trobe University report a rapid and scalable production of nanovesicles (NVs) directly from human trophectoderm cells (hTSCs) via serial mechanical extrusion of cells; these NVs can be generated in approximately 6 h with a 20-fold higher yield than EVs isolated from culture medium of the same number of cells. NVs display similar biophysical traits (morphologically intact, spherical, 90-130 nm) to EVs, and are laden with hallmark players of implantation that include cell-matrix adhesion and extracellular matrix organisation proteins (ITGA2/V, ITGB1, MFGE8) and antioxidative regulators (PRDX1, SOD2). Functionally, NVs are readily taken up by low-receptive endometrial HEC1A cells and reprogram their proteome towards a receptive phenotype that support hTSC spheroid attachment. Moreover, a single dose treatment with NVs significantly enhanced adhesion and spreading of mouse embryo trophoblast on fibronectin matrix. Thus, the researchers demonstrate the functional potential of NVs in enhancing embryo implantation and highlight their rapid and scalable generation, amenable to clinical utility.

Proteome profiling of HEC1A cells treated with NVs, EVs, or PBS

(A) Workflow for mass spectrometry-based proteomic profiling, comprising sera-bead sample preparation with nano-liquid chromatography tandem mass spectrometry and data processing/informatics. Created with Biorender. (B) Principal component analysis of HEC1A cellular proteome treated with NVs, EVs, or PBS control (volume matched). (C) Venn diagram of proteins identified in HEC1A cells after treatment with NV, EV, or PBS. (D) Two-way scatter plot highlighted the most significantly upregulated and downregulated proteins in NV-treated and EV-treated HEC1A cells compared to PBS control. (E) EnrichmentMap of Gene Ontology (biological process) and Reactome processes overrepresented in significantly upregulated proteins in both NV-treated and EV-treated HEC1A cells compared to PBS control.