Department of Laboratory Medicine, Karolinska Institutet, Huddinge, 14152, Sweden.
Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, Cairo University, Cairo, 11562, Egypt.
Adv Healthc Mater. 2022 Mar;11(5):e2101658. doi: 10.1002/adhm.202101658. Epub 2021 Nov 21.
Extracellular vesicles (EVs) are nanosized cell-derived vesicles produced by all cells, which provide a route of intercellular communication by transmitting biological cargo. While EVs offer promise as therapeutic agents, the molecular mechanisms of EV biogenesis are not yet fully elucidated, in part due to the concurrence of numerous interwoven pathways which give rise to heterogenous EV populations in vitro. The equilibrium between the EV-producing pathways is heavily influenced by factors in the extracellular environment, in such a way that can be taken advantage of to boost production of engineered EVs. In this study, a quantifiable EV-engineering approach is used to investigate how different cell media conditions alter EV production. The presence of serum, exogenous EVs, and other signaling factors in cell media alters EV production at the physical, molecular, and transcriptional levels. Further, it is demonstrated that the ceramide-dependent EV biogenesis route is the major pathway to production of engineered EVs during optimized EV-production. These findings suggest a novel understanding to the mechanisms underlying EV production in cell culture which can be applied to develop advanced EV production methods.
细胞外囊泡 (EVs) 是由所有细胞产生的纳米级细胞衍生囊泡，通过传递生物货物提供细胞间通讯途径。虽然 EVs 作为治疗剂具有很大的应用前景，但 EV 生物发生的分子机制尚未完全阐明，部分原因是存在许多相互交织的途径，导致体外产生异质 EV 群体。EV 产生途径之间的平衡受到细胞外环境中各种因素的严重影响，可以利用这些因素来促进工程 EV 的产生。在这项研究中，采用可量化的 EV 工程方法来研究不同的细胞培养基条件如何改变 EV 的产生。细胞培养基中血清、外源性 EV 和其他信号因子的存在会在物理、分子和转录水平上改变 EV 的产生。此外，还证明了在优化的 EV 产生过程中，依赖神经酰胺的 EV 生物发生途径是产生工程 EV 的主要途径。这些发现为细胞培养中 EV 产生的机制提供了新的认识，可用于开发先进的 EV 产生方法。
