Comparison of Cell and Organoid-Level Analysis of Patient-Derived 3D Organoids to Evaluate Tumor Cell Growth Dynamics and Drug Response
Autor: | Erin Spiller, Natasha Hashemi, Jasmine Foo, Roy Lau, Nolan Ung, Jordan Gasho, Ren X. Sun, Sarah Choung, Seung Il Kim, Emma J. Fong, Shannon M. Mumenthaler |
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Rok vydání: | 2020 |
Předmět: |
0301 basic medicine
Colorectal cancer Confocal Cell Computational biology Biology Irinotecan confocal imaging Biochemistry Analytical Chemistry Sphericity 03 medical and health sciences 3D cell culture Imaging Three-Dimensional 0302 clinical medicine image analysis Live cell imaging medicine Organoid Humans Anthracyclines drug screening 3D patient-derived tumor organoids Cell Proliferation Original Research Microscopy Confocal Dynamics (mechanics) Staurosporine medicine.disease Organoids 030104 developmental biology medicine.anatomical_structure 030220 oncology & carcinogenesis Molecular Medicine Fluorouracil Drug Screening Assays Antitumor Colorectal Neoplasms Biotechnology |
Zdroj: | Slas Discovery |
ISSN: | 2472-5552 |
Popis: | 3D cell culture models have been developed to better mimic the physiological environments that exist in human diseases. As such, these models are advantageous over traditional 2D cultures for screening drug compounds. However, the practicalities of transitioning from 2D to 3D drug treatment studies pose challenges with respect to analysis methods. Patient-derived tumor organoids (PDTOs) possess unique features given their heterogeneity in size, shape, and growth patterns. A detailed assessment of the length scale at which PDTOs should be evaluated (i.e., individual cell or organoid-level analysis) has not been done to our knowledge. Therefore, using dynamic confocal live cell imaging and data analysis methods we examined tumor cell growth rates and drug response behaviors in colorectal cancer (CRC) PDTOs. High-resolution imaging of H2B-GFP-labeled organoids with DRAQ7 vital dye permitted tracking of cellular changes, such as cell birth and death events, in individual organoids. From these same images, we measured morphological features of the 3D objects, including volume, sphericity, and ellipticity. Sphericity and ellipticity were used to evaluate intra- and interpatient tumor organoid heterogeneity. We found a strong correlation between organoid live cell number and volume. Linear growth rate calculations based on volume or live cell counts were used to determine differential responses to therapeutic interventions. We showed that this approach can detect different types of drug effects (cytotoxic vs cytostatic) in PDTO cultures. Overall, our imaging-based quantification workflow results in multiple parameters that can provide patient- and drug-specific information for screening applications. |
Databáze: | OpenAIRE |
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