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Semi-Automated Computational Assessment of Cancer Organoid Viability Using Rapid Live-Cell Microscopy
Joseph D Buehler1, Cylaina E Bird1,2, Milan R Savani3
1O'Donnell Brain Institute, University of Texas Southwestern Medical Center, Dallas, TX, USA.
Abstract:
The creation of patient-derived cancer organoids represents a key advance in preclinical modeling and has recently been applied to a variety of human solid tumor types. However, conventional methods used to assess in vivo tumor tissue treatment response are poorly suited for the evaluation of cancer organoids because they are time-intensive and involve tissue destruction. To address this issue, we established a suite of 3-dimensional patient-derived glioma organoids, treated them with chemoradiotherapy, stained organoids with non-toxic cell dyes, and imaged them using a rapid laser scanning confocal microscopy method termed "Apex Imaging." We then developed and tested a fragmentation algorithm to quantify heterogeneity in the topography of the organoids as a potential surrogate marker of viability. This algorithm, SSDquant, provides a 3-dimensional visual representation of the organoid surface and a numerical measurement of the sum-squared distance (SSD) from the derived mass center of the organoid. We tested whether SSD scores correlate with traditional immunohistochemistry-derived cell viability markers (cellularity and cleaved caspase 3 expression) and observed statistically significant associations between them using linear regression analysis. Our work describes a quantitative, non-invasive approach for the serial measurement of patient-derived cancer organoid viability, thus opening new avenues for the application of these models to studies of cancer biology and therapy.
Insights
We developed a non-invasive method to measure cancer organoid viability using Apex Imaging and SSDquant. This approach quantifies tumor treatment response in preclinical models, advancing cancer research.
Area of Science:
- Oncology
- Biotechnology
- Medical Imaging
Background:
- Patient-derived cancer organoids are valuable preclinical models.
- Traditional methods for assessing treatment response in organoids are destructive and time-consuming.
- A need exists for non-invasive, quantitative methods to evaluate organoid viability.
Purpose of the Study:
- To develop and validate a non-invasive method for assessing patient-derived cancer organoid viability.
- To establish a quantitative surrogate marker for tumor treatment response in organoid models.
- To enable serial measurements of organoid viability for improved preclinical studies.
Main Methods:
- Established 3D patient-derived glioma organoids.
- Treated organoids with chemoradiotherapy.
- Employed Apex Imaging (rapid laser scanning confocal microscopy) and a fragmentation algorithm (SSDquant) to quantify surface topography.
- Correlated SSD scores with immunohistochemistry markers (cellularity, cleaved caspase 3) using linear regression.
Main Results:
- Developed SSDquant, a novel algorithm to measure organoid surface heterogeneity.
- Demonstrated statistically significant correlations between SSD scores and traditional cell viability markers.
- Validated Apex Imaging as a rapid, non-destructive imaging technique for organoids.
Conclusions:
- SSDquant provides a quantitative, non-invasive measure of cancer organoid viability.
- This method facilitates serial assessment of treatment response in organoid models.
- The approach offers new possibilities for cancer biology and therapy studies using organoids.
