Institute of Biomedicine, University of Eastern Finland, Kuopio, Finland.
EV Group, Molecular and Integrative Biosciences Research Programme, Faculty of Biological and Environmental Sciences, University of Helsinki, Helsinki, Finland.
J Extracell Vesicles. 2022 Oct;11(10):e12273. doi: 10.1002/jev2.12273.
Recent advances in cell biology research regarding extracellular vesicles have highlighted an increasing demand to obtain 3D cell culture-derived EVs, because they are considered to more accurately represent EVs obtained in vivo. However, there is still a grave need for efficient and tunable methodologies to isolate EVs from 3D cell cultures. Using nanofibrillar cellulose (NFC) scaffold as a 3D cell culture matrix, we developed a pipeline of two different approaches for EV isolation from cancer spheroids. A batch method was created for delivering high EV yield at the end of the culture period, and a harvesting method was created to enable time-dependent collection of EVs to combine EV profiling with spheroid development. Both these methods were easy to set up, quick to perform, and they provided a high EV yield. When compared to scaffold-free 3D spheroid cultures on ultra-low affinity plates, the NFC method resulted in similar EV production/cell, but the NFC method was scalable and easier to perform resulting in high EV yields. In summary, we introduce here an NFC-based, innovative pipeline for acquiring EVs from 3D cancer spheroids, which can be tailored to support the needs of variable EV research objectives.
最近细胞生物学研究中关于细胞外囊泡的进展强调了对获得 3D 细胞培养衍生的 EV 的需求不断增加，因为它们被认为更能准确地代表体内获得的 EV。然而，仍然迫切需要高效和可调的方法从 3D 细胞培养物中分离 EV。我们使用纳米原纤维纤维素 (NFC) 支架作为 3D 细胞培养基质，开发了从癌细胞球体中分离 EV 的两种不同方法的流水线。创建了一种批量方法来在培养期末获得高 EV 产量，创建了一种收获方法来实现随时间收集 EV，以将 EV 分析与球体发育相结合。这两种方法都易于设置、快速执行，并提供了高 EV 产量。与超亲和性平板上无支架 3D 球体培养相比，NFC 方法产生的 EV 产量/细胞相似，但 NFC 方法具有可扩展性且更易于执行，从而产生高 EV 产量。总之，我们在这里介绍了一种基于 NFC 的创新流水线，用于从 3D 癌症球体中获取 EV，可以根据不同的 EV 研究目标进行调整。
