Abstract:
The growing understanding of the role of extracellular vesicles (EVs) in embryo-maternal communication has sparked considerable interest in their therapeutic potential within assisted reproductive technology, particularly in enhancing implantation success. However, the major obstacle remains the large-scale production of EVs, and there is still a gap in understanding how different culture systems affect the characteristics of the EVs. In the current study, trophoblast analogue human chorionic carcinoma cell line was cultivated in both conventional monolayer culture (2D) and as spheroids in suspension culture (3D) and how the cell growth environment affects the physical, biochemical and cellular signalling properties of EVs produced by them was studied. Interestingly, the 3D system was more active in secreting EVs compared to the 2D system, while no significant differences were observed in terms of morphology, size, and classical EV protein marker expression between EVs derived from the two culture systems. There were substantial differences in the proteomic cargo profile and cellular signalling potency of EVs derived from the two culture systems. Notably, 2D EVs were more potent in inducing a cellular response in endometrial epithelial cells (EECs) compared to 3D EVs. Therefore, it is essential to recognize that the biological activity of EVs depends not only on the cell of origin but also on the cellular microenvironment of the parent cell. In conclusion, caution is warranted when selecting an EV production platform, especially for assessing the functional and therapeutic potential of EVs through in vitro studies.
摘要:
对细胞外囊泡(EV)在胚胎-母体交流中的作用的了解日益增加,引起了人们对其在辅助生殖技术中的治疗潜力的极大兴趣。特别是在提高植入成功率。然而,主要障碍仍然是电动汽车的大规模生产,在理解不同的文化系统如何影响电动汽车的特征方面仍然存在差距。在目前的研究中,在常规单层培养(2D)和悬浮培养(3D)中培养滋养细胞类似物人绒毛膜癌细胞系,以及细胞生长环境如何影响物理,研究了它们产生的电动汽车的生化和细胞信号传导特性。有趣的是,与2D系统相比,3D系统在分泌电动汽车方面更加活跃,虽然在形态学方面没有观察到显著差异,尺寸,和来自两个培养系统的EV之间的经典EV蛋白标记表达。源自两种培养系统的EV的蛋白质组货物谱和细胞信号传导效力存在实质性差异。值得注意的是,与3DEV相比,2DEV在诱导子宫内膜上皮细胞(EEC)的细胞反应方面更有效。因此,必须认识到,电动汽车的生物活性不仅取决于起源细胞,还取决于亲本细胞的细胞微环境。总之,选择电动汽车生产平台时要谨慎,特别是通过体外研究评估电动汽车的功能和治疗潜力。