Speaker
Description
Our work involves the formation and utilization of mass produced, monodisperse spheroids encapsulated in hollow hydrogel microcapsules for use in assays and drug response trials.
Three-dimensional culture of cells into spheroids and organoids shows improved therapeutic and clinical relevance over two-dimensional culture. Heterogeneity of spheroids in size and structure is an issue for reproducibility, as such high-throughput, monodisperse production is the target of new culture methods.
Microfluidic Flow-Focusing is a high-throughput method for producing monodisperse droplets. By mixing cells in thermo-reversible hydrogel and producing a Water-In-Oil emulsion, cells can be encapsulated in spherical monodisperse hydrogel scaffolds. These scaffolds can then be re-encapsulated in a second layer of hydrogel of a higher melting temperature, to produce hollow hydrogel microcapsules with a tunable and monodisperse inner volume allowing growth of cells into spheroids of a maximum diameter.
Using this encapsulation method, roughly 100,000 of these cell laden microcapsules can be produced in an hour, with spheroids being fully formed after 5-7 days culture. These spheroids have a monodispersity of 16%, which is competitive with existing methods, and due to the containment of the hydrogel shell these sizes remained consistent for at least two weeks culture without any negative impact on viability.
Due to the robust hydrogel shell, these spheroids can be manipulated without risk of damage, allowing for easier partitioning of spheroids post culture or their insertion into microfluidic platforms.
Experiments have shown improved production of extracellular vesicles over cells cultured in well plates, showing promise for either paracrine therapies, or for studies that monitor changes in intercellular signalling pathways.
These encapsulated spheroids also show the typical dose related response to cancer drugs such as Paclitaxel and Doxorubicin, allowing for their use in drug discovery.
Ongoing work involves the formation of a microfluidic array device designed for the isolation of individual spheroids to allow for targeted dosing and imaging with replication. This array will also allow for reporter assays between two different spheroid types in a well, utilizing the robust isolation offered by the hydrogel shell to prevent direct contact while allowing vesicle transfer.
| Keyword-1 | Microfluidics |
|---|---|
| Keyword-2 | Drug Discovery |
| Keyword-3 | Hydrogel Scaffolds |