Using Single Cell Manipulation and Micropatterned Biomimetic Molecules to Build 2.5D/3D Neuronal Networks
Swiss partners
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ETH Zürich: Janos Vörös (main applicant), Christina Myra Tringides
Partners in the MENA region
- American University of Beirut, Liban: Rami Mhanna (main applicant)
Presentation of the projet
Injuries and diseases of the nervous tissues are among the most devastating conditions affecting humans and their treatment remains a major biomedical challenge. A great barrier for the development of therapies for neural injuries and diseases is our poor understanding of pathophysiological cues that govern these conditions, as well as the mechanisms by which information is stored and processed. This lack of knowledge is largely attributed to the difficulty of studying neural behavior in vivo because of the complexity of the tissue and challenging access of experimental tools (e.g. patch clamp and electrodes among others). In vitro cultures on the other hand mostly fail to mimic the controlled cell-cell contact and are largely based on random two dimensional (2D) neural networks.
Here we propose to create 2D and three dimensional (3D) neural networks by a combination of two direct solution deposition techniques; the first is deposit single cells from a novel bioprinting device (FluidFM® BOT, Cytosurge AG, Switzerland), and the second is using 3D printing to deposit hydrogel materials. Patterns of biomimetic extracellular matrix molecules (e.g. alginate sulfate) that favor the binding of neurons will be placed at pre-defined positions in the substrate where envisioned electrodes are present. Single neurons will then be picked and dropped exactly on the adhesion spots using the FluidFM® BOT bioprinter. Neural axons will be guided to grow along in situ drawn paths using lines with gradients of alginate sulfate that have embedded growth factors. Adsorption of the materials to the substrate will make use of biotinstreptavidin bonding resulting in stable patterns. To construct 3D networks, a thin biotinylated alginate hydrogel will be spincoated on the substrate. Conductive nanomaterials embedded in the matrix can give the material electrical conductivity and further promote the growth of neural cells. Patterns will then be performed in a similar way as above. Firing events will be measured using calcium imaging or with electrode arrays. Relevant protein deposition including axonal and dendritic markers will be detected by immunohistochemistry. The proposed research will bring forth a controlled neural network using a novel low-cost biomimetic molecule. Such systems can be used to gain better insight into the healthy development of the brain as well as changes during injuries and diseases.