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Monday, October 21 • 12:10pm - 12:30pm
Enabling Technologies- Integrated Droplet Microfluidic Platform for 3D Oncology Studies

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Spheroids are appealing candidates for replacing 2D cultures since they present a higher level of biological relevance and can be produced at high throughput. Their broad acceptance is hindered by the difficult adaptation to automated screening workflows that have been developed for conventional 2D systems.
In a previous work, we presented an integrated droplet microfluidic chip for the high throughput and reliable production of homogeneous spheroids, for their long term culture and multiscale analysis (Sart, S., Tomasi, R. F.-X., Amselem, G. & Baroud, C. N. Nat. Commun. (2017)). Fluorescent signals and morphological parameters of tens of thousands of spheroids and hundreds of thousands of individual cells were analyzed and correlated in situ. Spheroids were produced in droplets that were trapped in defined locations on chip.

This immobilization allowed easy imaging through time and any perfusion protocol to be performed on chip, like drug perfusion or in situ immuno-cyto-chemistry procedures (including washing steps). More recently we demonstrated the ability to add some content to the droplets at any given time during the experiment through controlled droplet pair merging for controlled 3D co-culture and drug toxicity screening on chip (Tomasi, R. F.-X., Sart, S., Champetier, T. & Baroud, C. N. bioRxiv, (2018)). First droplets were produced, immobilized, and kept in culture for spheroid formation, while second droplets were trapped and merged few days later to bring another cell type or different concentrations of a drug to the initial droplets.

In the present work, this technique is used to perform 3D oncology studies. As the role of the microenvironment is acknowledged to be crucial in tumor progression, we used the sequential microfluidic droplet trapping and merging to build complex 3D tumor cultures. For instance, PC3 cells (prostate cancer) were cultured in 3D with human mesenchymal stem cells that are known to be strong regulators of connective tissues. We are also investigating immunotherapy responses by putting into contact mouse melanoma spheroids (B16-F0) formed after 2 days of cultures and primary mouse cytotoxic T-cells. The microenvironment can also be mimicked by implementing extracellular matrix. For instance we built invasion assays in droplets: after formation of mouse melanoma spheroids in liquid droplets, Matrigel droplets were added and the resulting droplets gelified around each spheroid. One day later, the tumor cells began to invade the matrix.

As cancer progression is inherently associated to tumor growth, we performed long term culture of highly tumorigenic cells in the microfluidic chip. Ewing’s sarcoma spheroids (A673) grew from 60 to 160 µm in diameter within a week a culture in the droplets, each individual being monitored individually by imaging. Finally, the susceptibility of these spheroids to cis-platin was assessed over more than 4 decades of drug concentration on chip.

This technology integrates complex 3D culture protocols on a single microfluidic format, which greatly simplify workflows while benefiting from the miniaturization (higher density, cost savings, low sample volumes). By providing reproducible complex 3D environments at high throughput, this technology could have a strong impact in oncology, drug discovery and personalized medicine.

Aude DURAND, Pasteur
Aimee WESSEL, Pasteur
Gustave RONTEIX, Pasteur
Christelle ANGELY, Pasteu
Elaine DEL NERY, Curie
Charles BAROUD, Pasteur

avatar for Raphael Tomasi

Raphael Tomasi

Post Doctoral Researcher, Ecole Polytechnique Institut Pasteur
Raphaël TOMASI graduated from the ESPCI Paris in 2013 with an engineering degree as well as a research master in microfluidics. He gained his PhD from the hydrodynamics laboratory (LadHyX) of the Ecole Polytechnique (Paris area) in 2016, under the supervision of Charles BAROUD. Raphaël... Read More →

avatar for Hazel Screen, Ph.D., MRes, BEng

Hazel Screen, Ph.D., MRes, BEng

Professor of Biomedical Engineering, Queen Mary University of London
Hazel Screen is Professor of Biomedical Engineering and Chair of Bioengineering at Queen Mary University of London.Her research group focuses on multi-scale structure-function behaviour and mechanobiology in tissues. She has particular interest in in vitro models to explore health... Read More →

Monday October 21, 2019 12:10pm - 12:30pm BST
Wellcome Auditorium