Proteomic and Metabolomic Analyses of Adipose-derived Mesenchymal Stem Cell Exosomes and Culture Supernatants.

BACKGROUND/AIM: Aberrant differentiation of fertilized eggs during in vitro fertilization is a major contributor to infertility, and adipose-derived mesenchymal stem cells (ASCs) and their exosomes have been reported to facilitate fertilized egg differentiation; however, the underlying mechanisms remain unclear. This study aimed to elucidate how ASC-derived exosomes promote fertilized egg differentiation through proteomic analysis of ASC-derived exosomal proteins and metabolomic profiling of culture supernatants.

MATERIALS AND

METHODS

Extracellular vesicles were isolated from ASC culture supernatants via differential ultracentrifugation, proteins were extracted and digested using a phase-transfer surfactant protocol for LC-MS/MS analysis, and metabolites were analyzed using GC-MS/MS following extraction and derivatization.

RESULTS

Exosomes from early passage ASCs were enriched with various proteins, including alpha and beta proteasome subunits, which exhibit proteasome activity. Gene ontology analysis revealed the presence of proteins associated with transmembrane transport and cytoplasmic translation. Metabolomic profiling of culture supernatants revealed markedly elevated levels of amino acids associated with glycine, serine, and methionine metabolism.

CONCLUSION

ASC-derived culture supernatants can exert differentiation-promoting effects on fertilized eggs via exosomal proteins and soluble metabolites and may therefore be a novel option for infertility treatment.