A suspension technique for efficient large-scale cancer organoid culturing and perturbation screens.
Authors
Price, Stacey
Bhosle, Shriram
Gonçalves, Emanuel
Li, Xiaodun
McClurg, Dylan P
Barthorpe, Syd
Beck, Alex
Hall, Caitlin
Lightfoot, Howard
Farrow, Luke
Ansari, Rizwan
Jackson, David A
Allen, Laura
Roberts, Kirsty
Beaver, Charlotte
Francies, Hayley E
Garnett, Mathew J
Publication Date
2022-04-02Journal Title
Sci Rep
ISSN
2045-2322
Publisher
Springer Science and Business Media LLC
Volume
12
Issue
1
Language
en
Type
Article
This Version
VoR
Metadata
Show full item recordCitation
Price, S., Bhosle, S., Gonçalves, E., Li, X., McClurg, D. P., Barthorpe, S., Beck, A., et al. (2022). A suspension technique for efficient large-scale cancer organoid culturing and perturbation screens.. Sci Rep, 12 (1) https://doi.org/10.1038/s41598-022-09508-y
Abstract
Organoid cell culture methodologies are enabling the generation of cell models from healthy and diseased tissue. Patient-derived cancer organoids that recapitulate the genetic and histopathological diversity of patient tumours are being systematically generated, providing an opportunity to investigate new cancer biology and therapeutic approaches. The use of organoid cultures for many applications, including genetic and chemical perturbation screens, is limited due to the technical demands and cost associated with their handling and propagation. Here we report and benchmark a suspension culture technique for cancer organoids which allows for the expansion of models to tens of millions of cells with increased efficiency in comparison to standard organoid culturing protocols. Using whole-genome DNA and RNA sequencing analyses, as well as medium-throughput drug sensitivity testing and genome-wide CRISPR-Cas9 screening, we demonstrate that cancer organoids grown as a suspension culture are genetically and phenotypically similar to their counterparts grown in standard conditions. This culture technique simplifies organoid cell culture and extends the range of organoid applications, including for routine use in large-scale perturbation screens.
Keywords
Article, /631/80, /631/154, /631/208, /631/67, /631/67/69, /631/67/70, article
Sponsorship
Wellcome Trust (206194, 206194, 206194, 206194, 206194, 206194, 206194, 206194, 206194, 206194, 206194, 206194, 206194, 206194, 206194)
Medical Research Council (RG84369, RG84369)
Identifiers
s41598-022-09508-y, 9508
External DOI: https://doi.org/10.1038/s41598-022-09508-y
This record's URL: https://www.repository.cam.ac.uk/handle/1810/335708
Rights
Licence:
http://creativecommons.org/licenses/by/4.0/
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