Abstract
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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
van Norman GA (2020) Limitations of animal studies for predicting toxicity in clinical trials: part 2: potential alternatives to the use of animals in preclinical trials. JACC Basic Transl Sci 5:387–397
Ando Y, Ta HP, Yen DP, Lee SS, Raola S, Shen K (2017) A microdevice platform recapitulating hypoxic tumor microenvironments. Sci Rep 7:1–12
Eilenberger C, Rothbauer M, Selinger F, Gerhartl A, Jordan C, Harasek M, Schädl B, Grillari J, Weghuber J, Neuhaus W, Küpcü S, Ertl P (2021) A microfluidic multisize spheroid array for multiparametric screening of anticancer drugs and blood–brain barrier transport properties. Adv Sci (Weinh) 8:1–20
Refet-Mollof E, Najyb O, Chermat R, Glory A, Lafontaine J, Wong P, Gervais T (2021) Hypoxic jumbo spheroids on-A-chip (HOnAChip): insights into treatment efficacy. Cancers (Basel) 13:1–20
Moshksayan K, Kashaninejad N, Warkiani ME, Lock JG, Moghadas H, Firoozabadi B, Saidi MS, Nguyen NT (2018) Spheroids-on-a-chip: recent advances and design considerations in microfluidic platforms for spheroid formation and culture. Sensors Actuators B Chem 263:151–176
Bhattacharya S, Calar K, de La Puente P (2020) Mimicking tumor hypoxia and tumor-immune interactions employing three-dimensional in vitro models. J Exp Clin Cancer Res 39:1–16
Liu X, Fang J, Huang S, Wu X, Xie X, Wang J, Liu F, Zhang M, Peng Z, Hu N (2021) Tumor-on-a-chip: from bioinspired design to biomedical application. Microsyst Nanoeng 7:50
Wu Q, Liu J, Wang X, Feng L, Wu J, Zhu X, Wen W, Gong X (2020) Organ-on-a-chip: recent breakthroughs and future prospects. Biomed Eng Online 19:1–19
Marimuthu M, Kim S (2013) Pumpless steady-flow microfluidic chip for cell culture. Anal Biochem 437:161–163
Marimuthu M, Kim S (2015) Continuous oxygen supply in pump-less micro-bioreactor based on microfluidics. Biochip J 9:1–9
Marimuthu M, Rousset N, St-Georges-Robillard A, Lateef MA, Ferland M, Mes-Masson A-M, Gervais T (2018) Multi-size spheroid formation using microfluidic funnels. Lab Chip 18:304–314
Cavnar SP, Salomonsson E, Luker KE, Luker GD, Takayama S (2014) Transfer, imaging, and analysis plate for facile handling of 384 hanging drop 3D tissue spheroids. J Lab Autom 19:208–214
Acknowledgments
This work was supported by NSERC (RGPIN-06409), the Cancer Research Society, and CMC Microsystems (cmc.ca), the Fond de Recherche du Québec—Santé (FRQS) and FRQ-NT Quebec-India post-doctoral fellowship. M Marimuthu acknowledges the support of Dhanalakshmi Srinivasan University.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Marimuthu, M., Gervais, T. (2023). Multiplexed Viability Assays for High-Throughput Screening of Spheroids of Multiple Sizes. In: Friedrich, O., Gilbert, D.F. (eds) Cell Viability Assays. Methods in Molecular Biology, vol 2644. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3052-5_28
Download citation
DOI: https://doi.org/10.1007/978-1-0716-3052-5_28
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-3051-8
Online ISBN: 978-1-0716-3052-5
eBook Packages: Springer Protocols