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The Mini-Organo : A rapid high-throughput 3D coculture organotypic assay for oncology screening and drug development

Chitty, Jessica L. ; Skhinas, Joanna N. ; Filipe, Elysse C. ; Wang, Shan LU ; Cupello, Carmen Rodriguez LU orcid ; Grant, Rhiannon D. ; Yam, Michelle ; Papanicolaou, Michael ; Major, Gretel and Zaratzian, Anaiis , et al. (2020) In Cancer Reports 3(1).
Abstract
Background: The use of in vitro cell cultures is a powerful tool for obtaining key insights into the behaviour and response of cells to interventions in normal and disease situations. Unlike in vivo settings, in vitro experiments allow a fine-tuned control of a range of microenvironmental elements independently within an isolated setting. The recent expansion in the use of three-dimensional (3D) in vitro assays has created a number of representative tools to study cell behaviour in a more physiologically 3D relevant microenvironment. Complex 3D in vitro models that can recapitulate human tissue biology are essential for understanding the pathophysiology of disease. Aim: The development of the 3D coculture collagen contraction and invasion... (More)
Background: The use of in vitro cell cultures is a powerful tool for obtaining key insights into the behaviour and response of cells to interventions in normal and disease situations. Unlike in vivo settings, in vitro experiments allow a fine-tuned control of a range of microenvironmental elements independently within an isolated setting. The recent expansion in the use of three-dimensional (3D) in vitro assays has created a number of representative tools to study cell behaviour in a more physiologically 3D relevant microenvironment. Complex 3D in vitro models that can recapitulate human tissue biology are essential for understanding the pathophysiology of disease. Aim: The development of the 3D coculture collagen contraction and invasion assay, the "organotypic assay," has been widely adopted as a powerful approach to bridge the gap between standard two-dimensional tissue culture and in vivo mouse models. In the cancer setting, these assays can then be used to dissect how stromal cells, such as cancer-associated fibroblasts (CAFs), drive extracellular matrix (ECM) remodelling to alter cancer cell behaviour and response to intervention. However, to date, many of the published organotypic protocols are low-throughput, time-consuming (up to several weeks), and work-intensive with often limited scalability. Our aim was to develop a fast, high-throughput, scalable 3D organotypic assay for use in oncology screening and drug development. Methods and results Here, we describe a modified 96-well organotypic assay, the "Mini-Organo," which can be easily completed within 5 days. We demonstrate its application in a wide range of mouse and human cancer biology approaches including evaluation of stromal cell 3D ECM remodelling, 3D cancer cell invasion, and the assessment of efficacy of potential anticancer therapeutic targets. Furthermore, the organotypic assay described is highly amenable to customisation using different cell types under diverse experimental conditions. Conclusions: The Mini-Organo high-throughput 3D organotypic assay allows the rapid screening of potential cancer therapeutics in human and mouse models in a time-efficient manner. (Less)
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organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
3D model, cancer-associated fibroblast, coculture, drug screening, extracellular matrix, organotypic
in
Cancer Reports
volume
3
issue
1
article number
e1209
publisher
John Wiley & Sons Inc.
external identifiers
  • scopus:85085536506
  • pmid:32671954
ISSN
2573-8348
DOI
10.1002/cnr2.1209
language
English
LU publication?
yes
id
f1d61acb-1fbd-415b-911b-47ecd1da802d
date added to LUP
2019-09-12 16:40:50
date last changed
2022-05-04 00:51:12
@article{f1d61acb-1fbd-415b-911b-47ecd1da802d,
  abstract     = {{Background: The use of in vitro cell cultures is a powerful tool for obtaining key insights into the behaviour and response of cells to interventions in normal and disease situations. Unlike in vivo settings, in vitro experiments allow a fine-tuned control of a range of microenvironmental elements independently within an isolated setting. The recent expansion in the use of three-dimensional (3D) in vitro assays has created a number of representative tools to study cell behaviour in a more physiologically 3D relevant microenvironment. Complex 3D in vitro models that can recapitulate human tissue biology are essential for understanding the pathophysiology of disease. Aim: The development of the 3D coculture collagen contraction and invasion assay, the "organotypic assay," has been widely adopted as a powerful approach to bridge the gap between standard two-dimensional tissue culture and in vivo mouse models. In the cancer setting, these assays can then be used to dissect how stromal cells, such as cancer-associated fibroblasts (CAFs), drive extracellular matrix (ECM) remodelling to alter cancer cell behaviour and response to intervention. However, to date, many of the published organotypic protocols are low-throughput, time-consuming (up to several weeks), and work-intensive with often limited scalability. Our aim was to develop a fast, high-throughput, scalable 3D organotypic assay for use in oncology screening and drug development. Methods and results Here, we describe a modified 96-well organotypic assay, the "Mini-Organo," which can be easily completed within 5 days. We demonstrate its application in a wide range of mouse and human cancer biology approaches including evaluation of stromal cell 3D ECM remodelling, 3D cancer cell invasion, and the assessment of efficacy of potential anticancer therapeutic targets. Furthermore, the organotypic assay described is highly amenable to customisation using different cell types under diverse experimental conditions. Conclusions: The Mini-Organo high-throughput 3D organotypic assay allows the rapid screening of potential cancer therapeutics in human and mouse models in a time-efficient manner.}},
  author       = {{Chitty, Jessica L. and Skhinas, Joanna N. and Filipe, Elysse C. and Wang, Shan and Cupello, Carmen Rodriguez and Grant, Rhiannon D. and Yam, Michelle and Papanicolaou, Michael and Major, Gretel and Zaratzian, Anaiis and Da Silva, Andrew M. and Tayao, Michael and Vennin, Claire and Timpson, Paul and Madsen, Chris D. and Cox, Thomas R.}},
  issn         = {{2573-8348}},
  keywords     = {{3D model; cancer-associated fibroblast; coculture; drug screening; extracellular matrix; organotypic}},
  language     = {{eng}},
  number       = {{1}},
  publisher    = {{John Wiley & Sons Inc.}},
  series       = {{Cancer Reports}},
  title        = {{The Mini-Organo : A rapid high-throughput 3D coculture organotypic assay for oncology screening and drug development}},
  url          = {{http://dx.doi.org/10.1002/cnr2.1209}},
  doi          = {{10.1002/cnr2.1209}},
  volume       = {{3}},
  year         = {{2020}},
}