Abstract
Mesenchymal stem cells (MSCs) and MSC-derived extracellular vesicles (MSC EVs) have gained significant attention in biomedical and therapeutic applications. Nevertheless, their translation in clinical practice remains limited due to the lack of scalable manufacturing techniques and the prevailing safety concerns. Cell-derived extruded nanovesicles (eNVs) with high production efficiency are regarded as promising substitutes of EVs. However, like MSC EVs, their potential for tumorigenicity has yet to be exhaustively investigated. In this work, we investigated the tumorigenicity of umbilical cord mesenchymal stem cell-derived eNVs (UMSC eNVs). A549 cancer cell-derived eNVs (A549 eNVs) with potential tumorigenicity were also prepared for comparative analysis. Our characterization findings revealed that, although UMSC eNVs and A549 eNVs exhibited similar morphologies, they differed in their molecular composition. Subsequent animal experiments demonstrated the low tumorigenicity risk of UMSC eNVs in inducing tumor pathogenesis and development. Furthermore, microRNAs (miRNAs) profiling analyses suggested that the reduced tumorigenicity of UMSC eNVs might be due to the downregulation of hsa-miR-21-5p_R+1 and hsa-miR-192-5p, and upregulation of hsa-miR-143-3p and hsa-miR-146a-5p compared to A549 eNVs. The present study provided direct experimental confirmation and underlying miRNA profiling evidence of the biosafety of UMSC eNVs in terms of tumorigenicity, which will promote the future advancement and translation of UMSC eNVs.