Open AccessA quick pipeline for the isolation of 3D cell culture‐derived extracellular vesicles
Author(s) -
Kyykallio Heikki,
Faria Alessandra V. S.,
Hartman Rosabella,
Capra Janne,
Rilla Kirsi,
Siljander Pia RM
Publication year - 2022
Publication title -
journal of extracellular vesicles
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.94
H-Index - 68
ISSN - 2001-3078
DOI - 10.1002/jev2.12273
Subject(s) - 3d cell culture , spheroid , cell culture , computer science , extracellular vesicles , scaffold , pipeline (software) , scalability , microvesicles , computational biology , nanotechnology , microbiology and biotechnology , cell , chemistry , biological system , materials science , biology , microrna , biochemistry , operating system , database , gene , genetics
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.