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Advantages and limitations of using cell viability assays for 3D bioprinted constructs
Sofia Avnet1, Gemma Di Pompo2, Giorgia Borciani2
1Department of Biomedical and Neuromotor Sciences, Alma Mater Studiorum-Università di Bologna, Bologna, Italy.
Biomedical Materials (Bristol, England)
|February 2, 2024
Summary
Assessing cell viability in 3D bioprinted models is challenging. Automated confocal microscopy offers a more accurate, reproducible, and efficient method for evaluating cell viability and interactions in bioprinted tissues.
Area of Science:
- Biomaterials Science
- Tissue Engineering
- Cell Biology
Background:
- Bioprinting is crucial for creating bioengineered scaffolds and 3D disease models.
- Assessing the viability of cells within these complex 3D structures presents significant challenges.
- Traditional cell viability assays struggle with multi-layered bioink matrices and dispersed cells.
Purpose of the Study:
- To evaluate and compare different methods for assessing cell viability in bioprinted osteogenic constructs.
- To identify a more accurate, reproducible, and efficient technique for cell viability assessment in 3D bioprinting.
- To investigate the utility of automated confocal microscopy for analyzing cell distribution and interactions in bioprinted models.
Main Methods:
- Bioprinting of alginate- or gelatin-based bioinks loaded with osteogenic cells and ceramic microparticles.
- Comparison of manual cell counting, live/dead staining, alamarBlue assay, and automated confocal microscopy.
- Utilizing dual-photon confocal scanning for high-resolution imaging through the z-axis of 3D bioprints.
Main Results:
- Manual cell counting and live/dead staining showed high viability (80%-90%) but were time-consuming and operator-dependent.
- The alamarBlue assay was faster but less accurate due to hydrogel permeability issues.
- Automated confocal microscopy provided more reproducible, reliable, and faster results, avoiding overestimation compared to manual methods.
- Dual-photon confocal scanning enhanced precision in evaluating cell distribution, viability, and cell-cell interactions in thick 3D bioprints.
Conclusions:
- Automated confocal microscopy and cell counting significantly improve the accuracy of cell viability assessment in 3D bioprinted models.
- This advanced imaging technique offers superior precision for evaluating cell distribution and interactions compared to conventional methods.
- The findings highlight a more robust approach for quality control and research in the field of 3D bioprinting and tissue engineering.
