Measuring the viability of cancer spheroids
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Measuring the Viability of Cancer Spheroids
Introduction to Cancer Spheroids Viability
Cancer spheroids are three-dimensional (3D) cell culture models that closely mimic the in vivo tumor microenvironment. They are crucial for studying cancer biology and testing anticancer drugs. Measuring the viability of these spheroids is essential for evaluating the effectiveness of therapeutic agents and understanding tumor behavior.
Methods for Generating Cancer Spheroids
Hanging Drop Arrays for Ovarian Cancer Spheroids
A novel 384-well hanging drop array platform has been developed to generate stable, uniform ovarian cancer spheroids using as few as 10 cells. This method produces spheroids with high viability (over 85%) and demonstrates greater resistance to chemotherapy compared to 2D cultures.
Microfluidic Platforms for Pleural Cancer Spheroids
Microfluidic platforms allow for the loading, trapping, and chemoresistive testing of multicellular pleural cancer spheroids. This method uses caspase activity in the supernatant to detect spheroid viability, requiring minimal cell numbers, which is advantageous for personalized medicine applications.
Techniques for Measuring Viability
Electrical Impedance Tomography (EIT)
EIT is a label-free, real-time method for monitoring cell viability in large spheroids. It provides high spatial resolution and can detect cell mortality induced by chemicals, making it suitable for 3D drug screening and tissue engineering applications.
Lactate Dehydrogenase (LDH) Activity
LDH activity is used to quantify viability in organotypic multicellular spheroids of malignant glioma. This method involves staining cryostat sections and using automated image cytometry to measure LDH-active areas, providing reliable and reproducible results.
High Content Image Cytometry
The Celigo Image Cytometer measures real-time viability and apoptosis in 3D multicellular tumor spheroids using PI and caspase 3/7 stains. This method allows for the kinetic detection of drug effects, improving the identification of potential cancer drug candidates.
Electrochemical Detection
A hybrid gold nanostructure platform enables the formation of 3D cancer spheroids and the electrochemical detection of anticancer effects. This method allows for non-destructive, real-time pharmacokinetic analysis of drug candidates, providing precise and high-throughput screening capabilities.
Challenges and Considerations
Standardization and Reproducibility
Generating uniformly sized spheroids remains challenging, as different 3D culture methods produce heterogeneous populations. Standardizing protocols and assays is crucial for reliable readouts. Pre-selecting spheroids of homogeneous volume and shape can reduce data variability in cytotoxicity tests .
Physiological Relevance
3D spheroids better mimic the in vivo tumor environment compared to 2D cultures. They exhibit physico-chemical gradients, mechanical properties, and cell-cell interactions similar to those in tumors. These features are essential for accurate drug screening and understanding tumor biology .
Conclusion
Measuring the viability of cancer spheroids is vital for cancer research and drug development. Various methods, including EIT, LDH activity, high content image cytometry, and electrochemical detection, offer reliable and reproducible results. Standardizing spheroid generation and ensuring physiological relevance are key to obtaining meaningful data. These advancements in spheroid viability measurement pave the way for more effective cancer therapies and personalized medicine approaches.
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