Institute for Infection Prevention and Hospital Epidemiology, Medical Center-University of Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, Germany.
Institute for Infection Prevention and Hospital Epidemiology, Medical Center-University of Freiburg, Faculty of Medicine, University of Freiburg, Freiburg, Germany; Xiangya Hospital Affiliated To Central South University, Changsha, Hunan Province, PR China.
Methods Enzymol. 2020;645:209-230. doi: 10.1016/bs.mie.2020.09.005. Epub 2020 Oct 12.
Extracellular vesicles (EVs) play a pivotal role in cancer progression. However, the majority of functional studies performed so far relies on data acquired in traditional 2D cultures. Because the spatial architecture of tissue is decisive for the cell fate, new cell models to study EV functions in the 3D environment must approximate in vitro models to the physiological conditions. Several models were developed during the last years, which may be suitable to serve as 3D models to study EVs; among them are hydrogels, solid scaffolds, bioreactors, and 3D CoSeedis™ inserts. We present in this chapter a protocol for a 3D cell model based on the 3D CoSeedis™ agarose inserts, allowing for a long-term culture of cells of different origins under serum-free conditions and easy EV recovery. Additionally, information on individual culture conditions in 3D CoSeedis™ for different cell lines, protocols for model evaluation, and quality controls are included. We hope that our suggestions and experience will be useful to carry out EV study under more physiological conditions and contribute to the EV research field's progress.
细胞外囊泡（EVs）在癌症进展中起着关键作用。然而，迄今为止进行的大多数功能研究都依赖于在传统的 2D 培养中获得的数据。由于组织的空间结构对细胞命运具有决定性影响，因此，研究 EV 在 3D 环境中的功能的新细胞模型必须使体外模型更接近生理条件。近年来已经开发了几种模型，它们可能适合用作研究 EV 的 3D 模型；其中包括水凝胶、固体支架、生物反应器和 3D CoSeedis™插入物。我们在本章中提出了一种基于 3D CoSeedis™琼脂糖插入物的 3D 细胞模型的方案，允许在无血清条件下长期培养不同来源的细胞，并可轻松回收 EV。此外，还包括有关不同细胞系在 3D CoSeedis™中的单个培养条件的信息、模型评估的方案以及质量控制。我们希望我们的建议和经验将有助于在更接近生理的条件下进行 EV 研究，并为 EV 研究领域的发展做出贡献。
