Document Type
Article
Publication Date
9-6-2024
Original Citation
Bozal S,
Sjogren G,
Costa A,
Brown J,
Roberts S,
Baker D,
Gabriel P,
Ristau B,
Samuels M,
Flynn W,
Robson P,
Courtois E.
Development of an automated 3D high content cell screening platform for organoid phenotyping. SLAS Discov. 2024;29(7):100182.
JAX Source
SLAS Discov. 2024;29(7):100182.
ISSN
2472-5560
PMID
39245180
DOI
https://doi.org/10.1016/j.slasd.2024.100182
Grant
This work was supported by the National Cancer Institute of the National Institutes of Health under award numbers U01CA224013 and P30CA034196 and by the Scientific Services Innovation Fund of The Jackson Laboratory under award number 19005–18–01.
Abstract
The use of organoid models in biomedical research has grown substantially since their inception. As they gain popularity among scientists seeking more complex and biologically relevant systems, there is a direct need to expand and clarify potential uses of such systems in diverse experimental contexts. Herein we outline a high-content screening (HCS) platform that allows researchers to screen drugs or other compounds against three-dimensional (3D) cell culture systems in a multi-well format (384-well). Furthermore, we compare the quality of robotic liquid handling with manual pipetting and characterize and contrast the phenotypic effects detected by confocal imaging and biochemical assays in response to drug treatment. We show that robotic liquid handling is more consistent and amendable to high throughput experimental designs when compared to manual pipetting due to improved precision and automated randomization capabilities. We also show that image-based techniques are more sensitive to detecting phenotypic changes within organoid cultures than traditional biochemical assays that evaluate cell viability, supporting their integration into organoid screening workflows. Finally, we highlight the enhanced capabilities of confocal imaging in this organoid screening platform as they relate to discerning organoid drug responses in single-well co-cultures of organoids derived from primary human biopsies and patient-derived xenograft (PDX) models. Altogether, this platform enables automated, imaging-based HCS of 3D cellular models in a non-destructive manner, opening the path to complementary analysis through integrated downstream methods.
Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.