Method to Disassemble Spheroids into Core and Rim for Downstream Applications Such as Flow Cytometry, Comet Assay, Transcriptomics, Proteomics, and Lipidomics

Helle Sedighi Frandsen, Martina Štampar, Joel Mario Vej-Nielsen, Bojana Žegura, Adelina Rogowska-Wrzesinska*

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Abstrakt

Cells cultured in a monolayer have been a central tool in molecular and cell biology, toxicology, biochemistry, and so on. Therefore, most methods for adherent cells in cell biology are tailored to this format of cell culturing. Limitations and disadvantages of monolayer cultures, however, have resulted in the ongoing development of advanced cell culturing techniques. One such technique is culturing cells as multicellular spheroids, that had been shown to mimic the physiological conditions found in vivo more accurately. This chapter presents a novel method for separation of the spheroid rim and core in mature spheroids (>21 days) for further analysis using advanced molecular biology techniques such as flow cytometry, viability estimations, comet assay, transcriptomics, proteomics and lipidomic. This fast and gentle disassembly of intact spheroids into rim and core fractions, and further into viable single-cell suspension provides an opportunity to bridge the gap from 3D cell culture to current state-of-the-art analysis methods.

OriginalsprogEngelsk
TitelNext Generation Culture Platforms for Reliable In Vitro Models : Methods and Protocols
RedaktørerTiziana A.L. Brevini, Alireza Fazeli, Kursad Turksen
ForlagHumana Press
Publikationsdato2021
Sider173-188
ISBN (Trykt)978-1-0716-1245-3, Softcover ISBN 978-1-0716-1248-4
ISBN (Elektronisk) 978-1-0716-1246-0
DOI
StatusUdgivet - 2021
NavnMethods in Molecular Biology
Vol/bind2273
ISSN1064-3745

Bibliografisk note

Publisher Copyright:
© 2021, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

Copyright:
Copyright 2021 Elsevier B.V., All rights reserved.

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