Multiplexed live-cell imaging for drug responses in patient-derived organoid models of cancer

Kaitriana E Colling; Emily L Symons; Lorenzo Buroni; Hiruni K Sumanisiri; Jessica Andrew-Udoh; Emily Witt; Haley A Losh; Abigail M Morrison; Kimberly K Leslie; Christopher J Dunnill; Johann S De Bono; Kristina W Thiel

doi:10.1101/2023.11.15.567243

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Multiplexed live-cell imaging for drug responses in patient-derived organoid models of cancer

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Multiplexed live-cell imaging for drug responses in patient-derived organoid models of cancer

Kaitriana E Colling, Emily L Symons, Lorenzo Buroni, Hiruni K Sumanisiri, Jessica Andrew-Udoh, Emily Witt, Haley A Losh, Abigail M Morrison, Kimberly K Leslie, Christopher J Dunnill, …

bioRxiv : the preprint server for biology

Cold Spring Harbor Laboratory

11/17/2023

DOI: 10.1101/2023.11.15.567243

PMCID: PMC10680710

PMID: 38014133

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https://doi.org/10.1101/2023.11.15.567243View

Preprint (Author's original)This preprint has not been evaluated by subject experts through peer review. Preprints may undergo extensive changes and/or become peer-reviewed journal articles. Open Access

Abstract

Patient-derived organoid (PDO) models of cancer are a multifunctional research system that better recapitulates human disease as compared to cancer cell lines. PDO models can be generated by culturing patient tumor cells in extracellular basement membrane extracts (BME) and plating as three-dimensional domes. However, commercially available reagents that have been optimized for phenotypic assays in monolayer cultures often are not compatible with BME. Herein we describe a method to plate PDO models and assess drug effects using an automated live-cell imaging system. In addition, we apply fluorescent dyes that are compatible with kinetic measurements to simultaneously quantitate cell health and apoptosis. Image capture can be customized to occur at regular time intervals over several days. Users can analyze drug effects in individual Z-plane images or a Z Projection of serial images from multiple focal planes. Using masking, specific parameters of interest are calculated, such as PDO number, area, and fluorescence intensity. We provide proof-of-concept data demonstrating the effect of cytotoxic agents on cell health, apoptosis and viability. This automated kinetic imaging platform can be expanded to other phenotypic readouts to understand diverse therapeutic effects in PDO models of cancer.

Details

Title: Subtitle: Multiplexed live-cell imaging for drug responses in patient-derived organoid models of cancer
Creators: Kaitriana E Colling - Cancer Biology Graduate Program, Carver College of Medicine, University of Iowa, Iowa City, IA, USA
Emily L Symons - University of Iowa
Lorenzo Buroni - Royal Marsden NHS Foundation Trust
Hiruni K Sumanisiri - Department of Obstetrics and Gynecology, Carver College of Medicine, University of Iowa, Iowa City, IA, USA
Jessica Andrew-Udoh - University of Iowa
Emily Witt - Department of Radiation Oncology, Carver College of Medicine, University of Iowa, Iowa City, IA, USA
Haley A Losh - University of Iowa
Abigail M Morrison - University of Iowa
Kimberly K Leslie - University of New Mexico
Christopher J Dunnill - Agilent Technologies (United Kingdom)
Johann S De Bono - Royal Marsden NHS Foundation Trust
Kristina W Thiel - Holden Comprehensive Cancer Center, University of Iowa, Iowa City, IA, USA
Resource Type: Preprint
Publication Details: bioRxiv : the preprint server for biology
DOI: 10.1101/2023.11.15.567243
PMID: 38014133
PMCID: PMC10680710
Publisher: Cold Spring Harbor Laboratory; United States
Grant note: K22 CA263783 / NCI NIH HHS
Language: English
Date posted: 11/17/2023
Academic Unit: Radiation Oncology; Obstetrics and Gynecology
Record Identifier: 9984556659802771

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