Multiplexed Live-Cell Imaging for Drug Responses in Patient-Derived Organoid Models of Cancer

Overview

Journal J Vis Exp

Specialties Biology
General Medicine

Date 2024 Jan 22

PMID 38251777

Authors

Kaitriana E Colling

Emily L Symons

Lorenzo Buroni

Hiruni K Sumanasiri

Jessica Andrew-Udoh

Emily Witt

Haley A Losh

Abigail M Morrison

Kimberly K Leslie

Christopher J Dunnill

Johann S de Bono

Kristina W Thiel

Affiliations

Soon will be listed here.

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 them 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 quantify cell health and apoptosis simultaneously. 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.

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