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Extracellular vesicles (EVs) have been implicated in embryo\u2010maternal crosstalk, capable of reprogramming endometrial cells towards a pro\u2010implantation signature and phenotype. However, challenges associated with EV yield and direct loading of biomolecules limit their therapeutic potential. We have previously established generation of cell\u2010derived nanovesicles (NVs) from human trophectodermal cells (hTSCs) and their capacity to reprogram endometrial cells to enhance adhesion and blastocyst outgrowth. Here, we employed a rapid NV loading strategy to encapsulate potent implantation molecules such as HB\u2010EGF (NV<jats:sup>HBEGF<\/jats:sup>). We show these loaded NVs elicit EGFR\u2010mediated effects in recipient endometrial cells, activating kinase phosphorylation sites that modulate their activity (AKT S124\/129, MAPK1 T185\/Y187), and downstream signalling pathways and processes (AKT signal transduction, GTPase activity). Importantly, they enhanced target cell attachment and invasion. The phosphoproteomics and proteomics approach highlight NV<jats:sup>HBEGF<\/jats:sup>\u2010mediated short\u2010term signalling patterns and long\u2010term reprogramming capabilities on endometrial cells which functionally enhance trophectodermal\u2010endometrial interactions. This proof\u2010of\u2010concept study demonstrates feasibility in enhancing the functional potency of NVs in the context of embryo implantation.<\/jats:p>","DOI":"10.1002\/pmic.202200145","type":"journal-article","created":{"date-parts":[[2024,1,12]],"date-time":"2024-01-12T10:34:55Z","timestamp":1705055695000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["HB\u2010EGF\u2010loaded nanovesicles enhance trophectodermal spheroid attachment and invasion"],"prefix":"10.1002","volume":"24","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2620-2834","authenticated-orcid":false,"given":"Qi Hui","family":"Poh","sequence":"first","affiliation":[{"name":"Molecular Proteomics Baker Heart and Diabetes Institute  Melbourne Victoria Australia"},{"name":"Department of Biochemistry and Chemistry School of Agriculture Biomedicine and Environment La Trobe University  Bundoora Victoria Australia"},{"name":"Department of 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