Abstract：
Extracellular vesicles are secreted by all eukaryotic cells and they have an important role in intercellular signaling. Plant extracellular vesicles (PEVs) are a novel area of research that has gained attention due to their potential implications in biomolecule transport and therapeutic applications. PEVs are lipid bilayer-enclosed structures that contain a diverse cargo of biomolecules such as proteins and lipids. Moreover, it is known that PEVs have a noticeable therapeutic potential for various conditions such as inflammation and oxidative stress. However, there are critical problems such as removing the endosomes and plant-derived biomolecules that decrease the standardization and therapeutic efficacy of PEVs. In our study, the aim was to characterize plant cell suspension-derived extracellular vesicles (PCSEVs) obtained from two different plant cell suspension cultures: Stevia rebaudiana and Vaccaria hispanica. These vesicles were isolated using ultrafiltration and characterized with nanoparticle tracking analysis (NTA) and atomic force microscopy (AFM). The molecular composition of PCSEVs was profiled and the cellular uptake assay was performed. Our results demonstrated that PCSEVs have a spherical shape, less than 200 nm. In the fatty acid analysis, the primary components in PCSEVs were palmitic acid, linoleic acid, and cis-vaccenic acid. The protein content of Stevia rebaudiana-derived EVs (SDEVs) was largely associated with proteins involved in extracellular structures and functions. Conversely, Vaccaria hispanica-derived EVs (HDEVs) displayed a higher presence of cytosolic proteins. These findings contribute to the understanding of PCSEVs and open up potential avenues in extracellular vesicle research, pointing to promising prospects for future innovations in various fields.
摘要：
细胞外囊泡由所有真核细胞分泌，它们在细胞间信号传导中起重要作用。植物细胞外囊泡（PEV）是一个新的研究领域，由于其在生物分子转运和治疗应用中的潜在意义而受到关注。PEV是脂质双层封闭结构,其含有多种生物分子如蛋白质和脂质。此外，已知PEV对于各种病症如炎症和氧化应激具有显著的治疗潜力。然而,存在一些关键问题,例如去除核内体和植物衍生的生物分子,这些问题降低了PEV的标准化和治疗功效。在我们的研究中,目的是表征从两种不同的植物细胞悬浮培养物中获得的植物细胞悬浮衍生的细胞外囊泡（PCSEV）：甜叶菊rebaudiana和Vaccariahispanica。使用超滤分离这些囊泡，并用纳米粒子跟踪分析（NTA）和原子力显微镜（AFM）进行表征。分析PCSEV的分子组成并进行细胞摄取测定。我们的结果表明，PCSEV具有球形，小于200nm。在脂肪酸分析中，PCSEV的主要成分是棕榈酸，亚油酸,和顺式疫苗酸。甜叶菊衍生的电动汽车（SDEV）的蛋白质含量在很大程度上与细胞外结构和功能相关。相反，Vaccariamispanica衍生的EV（HDEV）显示出较高的胞浆蛋白存在。这些发现有助于对PCSEV的理解，并为细胞外囊泡研究开辟了潜在的途径。指出了各个领域未来创新的前景。
