Exosomes, nanosized vesicles secreted by cells, are gaining attention in biomedical research due to their biocompatibility, cargo-loading capacity, and ability to penetrate deep tissues. They serve as natural signaling agents in intercellular communication and can carry proteins, lipids, and nucleic acids, offering significant therapeutic potential. Exosomes can be used for various therapeutic applications, including chemotherapy, gene therapy, and photothermal therapy. Their homotypic targeting and self-recognition capabilities make them promising for personalized medicine. However, challenges remain in optimizing cargo loading efficiency, structural stability, and defining exosome origins. Future research should focus on developing large-scale, quality-controlled production methods, refining drug loading strategies, and conducting extensive in vivo studies and clinical trials. Exosomes are considered an interesting area in biomedical research due to their efficiency, stability, and safety as therapeutic delivery systems. Exosomes are the smallest type of extracellular vesicles, typically 30–150 nm in diameter, and originate from the endosomal system. They contain various molecular constituents, including proteins and RNA, from their cells of origin. Exosomes can be taken up by neighboring or distant cells and influence their function. Microvesicles, larger than exosomes, are formed by direct outward budding and fission of the plasma membrane. Apoptotic bodies are the largest type of EVs, generated during apoptosis. Exosomes and microvesicles are difficult to separate based solely on size, and no specific biomarkers distinguish them. The International Society for Extracellular Vesicles recommends classifying EVs into small EVs (sEVs, less than 200 nm) and medium/large EVs (m/lEVs, greater than 200 nm). The biogenesis of exosomes involves the endosomal pathway, starting with inward budding of the plasma membrane, leading to the formation of early endosomes, which mature into late endosomes. Late endosomes undergo inward budding of the multivesicular body (MVB) membrane, forming intraluminal vesicles (ILVs). These ILVs can fuse with the cellular membrane, releasing into the extracellular space. Alternatively, ILVs can merge with the lysosome if their contents are destined for degradation. Exosomes can be taken up by recipient cells through endocytosis, direct fusion with the plasma membrane, or interaction with cell-surface receptors, initiating intracellular signaling pathways. Endocytosis is the predominant mechanism for exosome uptake, but direct fusion is the most effective for delivering cargo into the cell. Exosomal molecular cargoes need to be released from vesicles into the cytoplasm, a process known as endosomal escape. If cargoes delivered via the endocytosis pathway remain within endosomes, they will be degraded by a lysosome or secreted into the extracellular space without therapeutic effect. Various methods have been developed to load therapeutic agents into exosomes, including passive loading andExosomes, nanosized vesicles secreted by cells, are gaining attention in biomedical research due to their biocompatibility, cargo-loading capacity, and ability to penetrate deep tissues. They serve as natural signaling agents in intercellular communication and can carry proteins, lipids, and nucleic acids, offering significant therapeutic potential. Exosomes can be used for various therapeutic applications, including chemotherapy, gene therapy, and photothermal therapy. Their homotypic targeting and self-recognition capabilities make them promising for personalized medicine. However, challenges remain in optimizing cargo loading efficiency, structural stability, and defining exosome origins. Future research should focus on developing large-scale, quality-controlled production methods, refining drug loading strategies, and conducting extensive in vivo studies and clinical trials. Exosomes are considered an interesting area in biomedical research due to their efficiency, stability, and safety as therapeutic delivery systems. Exosomes are the smallest type of extracellular vesicles, typically 30–150 nm in diameter, and originate from the endosomal system. They contain various molecular constituents, including proteins and RNA, from their cells of origin. Exosomes can be taken up by neighboring or distant cells and influence their function. Microvesicles, larger than exosomes, are formed by direct outward budding and fission of the plasma membrane. Apoptotic bodies are the largest type of EVs, generated during apoptosis. Exosomes and microvesicles are difficult to separate based solely on size, and no specific biomarkers distinguish them. The International Society for Extracellular Vesicles recommends classifying EVs into small EVs (sEVs, less than 200 nm) and medium/large EVs (m/lEVs, greater than 200 nm). The biogenesis of exosomes involves the endosomal pathway, starting with inward budding of the plasma membrane, leading to the formation of early endosomes, which mature into late endosomes. Late endosomes undergo inward budding of the multivesicular body (MVB) membrane, forming intraluminal vesicles (ILVs). These ILVs can fuse with the cellular membrane, releasing into the extracellular space. Alternatively, ILVs can merge with the lysosome if their contents are destined for degradation. Exosomes can be taken up by recipient cells through endocytosis, direct fusion with the plasma membrane, or interaction with cell-surface receptors, initiating intracellular signaling pathways. Endocytosis is the predominant mechanism for exosome uptake, but direct fusion is the most effective for delivering cargo into the cell. Exosomal molecular cargoes need to be released from vesicles into the cytoplasm, a process known as endosomal escape. If cargoes delivered via the endocytosis pathway remain within endosomes, they will be degraded by a lysosome or secreted into the extracellular space without therapeutic effect. Various methods have been developed to load therapeutic agents into exosomes, including passive loading and