Multiplexed Viability Assays for High-Throughput Screening of Spheroids of Multiple Sizes.
| Autor: | Marimuthu M; Department of Biomedical Engineering, Centre for Research, Dhanalakshmi Srinivasan University, Samayapuram, Trichy, Tamil Nadu, India., Gervais T; Department of Engineering Physics, Polytechnique Montréal, Chemin de Polytechnique, Montréal, QC, Canada. thomas.gervais@polymtl.ca.; Centre de Recherche du Centre Hospitalier de l'Université de Montréal, Montreal, QC, Canada. thomas.gervais@polymtl.ca. |
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| Jazyk: | angličtina |
| Zdroj: | Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2023; Vol. 2644, pp. 435-447. |
| DOI: | 10.1007/978-1-0716-3052-5_28 |
| Abstrakt: | High-throughput (HT) drug screening is in high demand for successful drug discovery and personalized medicine. Spheroids act as a promising preclinical model for HT drug screening, which may decrease drug failures in clinical trials. Numerous spheroid-forming technological platforms are currently under development, which include synchronous, jumbo-sized, hanging drop, rotary, and nonadherent surface spheroid growth. Initial cell seeding concentration and time of culture play a vital role for spheroids to mimic the extracellular microenvironment of natural tissue, especially for HT preclinical evaluation. Hence microfluidic platforms become a potential technology to provide a confined space for the oxygen and nutrient gradients within the tissues while controlling the cell count and spheroid size in an HT manner. We describe here a microfluidic platform capable of generating spheroids of multiple sizes in a controlled manner with a predefined cell concentration for HT drug screening. Ovarian cancer spheroids grown on this microfluidic platform were evaluated for viability using a confocal microscope and flow cytometer. In addition, screening of the HT chemotherapeutic drug carboplatin was carried out on-chip to evaluate the impact of spheroid size on drug toxicity. This chapter summarizes a detailed protocol on microfluidic platform fabrication for spheroid growth, on-chip multi-sized spheroid analysis, and chemotherapeutic drug screening. (© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.) |
| Databáze: | MEDLINE |
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