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Microfluidic device for high-throughput three-dimensional culture, mechanical stimulation and drug screening of stem cells

Final Report Summary - STEMCELLSTRESSCHIP (Microfluidic device for high-throughput three-dimensional culture, mechanical stimulation and drug screening of stem cells)

This fellowship was set out with the intention of developing a microfluidic platform to enable three-dimensional culture of stem cells within a high-throughput format. To achieve this, we employed our expertise in microfabrication and microfluidics and established novel protocols to fabricate networks of microfluidic channels on flexible substrates suitable for mechanical stimulation. Moreover, we also created advanced microfluidic devices compatible with automated cell culture and imaging that enabled highly parallel experiments with minimal quantities of reagents and cells, and allowed the dynamic study of intercellular processes in deep detail. We used the developed platforms to culture cells in controlled two- and three-dimensional microenvironments, and also to evaluate the behaviour of different cell types, including adult muscle stem cells, when varying the adhesion properties of the culture substrate as well as the composition of the culture medium.

We observed a clear response in cell morphology to the confinement in the microchannel environment, both in the case of culturing cells within an extracellular matrix as well as when seeding cells on the surface of the channels. To study this effect in detail, we created a microdevice with an array of channels of varying widths. We included active control modules that allowed us to perfuse cells regularly without shear flow, enabling the study of cell behaviour on low adhesion substrates. Finally, active flow control also allowed us to multiplex culture conditions and vary those combinatorially with factors such as cell type, extracellular matrix, drug concentration or adhesion ligands.

Stem cells are cells that have the ability to self-renew as well as to differentiate into other specific cell types. They have become one of the most active fields of biological research due to their importance in cancer, tissue homeostasis and regeneration, and their potential applications in tissue engineering and treatment of degenerative diseases. The technologies developed in this work will prove of great interest to the large biological community due to the impact they can have in the costly and difficult task of controlling the microenvironment of the stem cell. Therefore, the research carried out within this fellowship will be of direct interest not only to the scientific community, but also to the medical one and to society at large.