Comparative proteomic analysis of rat adipose-derived mesenchymal stem cells and their exosomes

Zhang Xinrui; Han Yue; Lei Lei; Liu Jianyu; Geng Chengkui

doi:10.12307/2025.046

Abstract：BACKGROUND:Research on adipose-derived mesenchymal stem cells and their exosomes often uses proteomics to analyze their roles in tissue repair,but comparative proteomic analyses between the two are scarce. OBJECTIVE:To analyze adipose-derived mesenchymal stem cells and their exosomes using proteomic analysis. METHODS:Rat adipose-derived mesenchymal stem cells were isolated,cultured,and identified.Exosomes were then extracted from cell supernatant and identified.Differentially expressed proteins of adipose-derived mesenchymal stem cells and their exosomes were analyzed using DIA proteomics.Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses were performed on these differentially expressed proteins. RESULTS AND CONCLUSION:(1) Adipose-derived mesenchymal stem cells displayed a mainly spindle-shaped,fibroblast-like morphology.(2) The exosome suspension protein concentration was 7.66 mg/mL,as determined by the BCA method.(3) Exosomes exhibited a characteristic teacup shape with a visible double-layer membrane vesicle structure.The center presented a low electron density component.The exosomes showed a peak particle size distribution of 112.2 nm,a concentration of 7.5×1011 particles/mL,and an average diameter ranging from 70 to 140 nm.(4) Exosomes expressed high levels of surface marker proteins CD9 and TSG101.(5) Gene Ontology analysis of differentially expressed proteins revealed enrichment in the extracellular matrix and synapses,with functions related to ion binding,ribosome binding,and particularly cell adhesion and translation.(6) Kyoto Encyclopedia of Genes and Genomes pathway analysis indicated that the differentially expressed proteins were primarily involved in extracellular matrix receptor interaction,ribosome,and cytokine receptor interaction,and also associated with various metabolic diseases like cholesterol and thyroid disorders.