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BACKGROUND: Extracellular vesicles (EVs) have attracted expanded attention as vehicles for the diagnosis and therapy of diseases and regenerative medicine due to their biocompatibility, efficient cellular uptake ability, and capacity to transport biologically active molecules. However, the low secretion yield of EVs and the challenges of large-scale production remain the main barriers to their extensive clinical use. METHODS AND RESULTS: This review explores recent strategies to enhance EV production in cell culture systems, focusing on chemical stimulation, mechanical stimulation, and structural stimulation. First, we review chemical stimulation strategies for modulating culture conditions using chemical stimulation, including nutrient composition, pH, temperature, oxygen levels, intracellular cholesterol, and oxidative stress. Second, we examine mechanical stimulation strategies, including shear stress, irradiation, and ultrasound. Third, we explore structural stimulation strategies, such as three-dimensional (3D) culture systems involving spheroid-based culture, as well as the use of bioreactors and scaffolds. In addition, cell-derived nanovesicles containing cell membrane and cellular component, which can be more easily mass-produced compared to EVs, are proposed as an alternative to EVs. CONCLUSION: Future research should focus on developing cost-effective and scalable EV production methods while improving purification techniques to ensure a high yield without compromising functional integrity. Moreover, integrating optimized stimulation strategies-such as refining 3D culture systems, bioreactor designs, and mechanical stimulation methods-could further enhance EV secretion. Addressing these challenges is essential for advancing EV-based applications in both research and clinical practice.
Park et al. (Thu,) studied this question.
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