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.

Details

Title
A suspension technique for efficient large-scale cancer organoid culturing and perturbation screens
Author
Price, Stacey 1 ; Bhosle Shriram 1 ; Gonçalves Emanuel 2 ; Li, Xiaodun 3 ; McClurg, Dylan P 3 ; Barthorpe Syd 1 ; Beck, Alex 1 ; Hall, Caitlin 1 ; Lightfoot, Howard 1 ; Farrow, Luke 1 ; Ansari Rizwan 1 ; Jackson, David A 1 ; Allen, Laura 1 ; Roberts, Kirsty 1 ; Beaver, Charlotte 1 ; Francies, Hayley E 1 ; Garnett, Mathew J 1 

 Wellcome Sanger Institute, Cambridge, UK (GRID:grid.10306.34) (ISNI:0000 0004 0606 5382) 
 Wellcome Sanger Institute, Cambridge, UK (GRID:grid.10306.34) (ISNI:0000 0004 0606 5382); Universidade de Lisboa, Instituto Superior Técnico (IST), Lisbon, Portugal (GRID:grid.9983.b) (ISNI:0000 0001 2181 4263); INESC-ID, Lisbon, Portugal (GRID:grid.14647.30) (ISNI:0000 0001 0279 8114) 
 University of Cambridge, MRC Cancer Unit, Cambridge, UK (GRID:grid.5335.0) (ISNI:0000000121885934) 
Publication year
2022
Publication date
2022
Publisher
Nature Publishing Group
e-ISSN
20452322
Source type
Scholarly Journal
Language of publication
English
ProQuest document ID
2646390559
Copyright
© The Author(s) 2022. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.