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A suspension technique for efficient large-scale cancer organoid culturing and perturbation screens.

Published version
Peer-reviewed

Change log

Authors

Price, Stacey 
Bhosle, Shriram 
Gonçalves, Emanuel 
Li, Xiaodun 
McClurg, Dylan P 

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.

Description

Keywords

Cell Culture Techniques, DNA, Humans, Neoplasms, Organoids

Journal Title

Sci Rep

Conference Name

Journal ISSN

2045-2322
2045-2322

Volume Title

12

Publisher

Springer Science and Business Media LLC
Sponsorship
Wellcome Trust (206194, 206194, 206194, 206194, 206194, 206194, 206194, 206194, 206194, 206194, 206194, 206194, 206194, 206194, 206194)
Medical Research Council (RG84369, RG84369)