Third party funded individual grant
Start date : 01.01.2013
End date : 31.12.2014
In many chronic degenerative and inflammatory brain diseases (e.g. multiple sclerosis, Alzheimer's Disease), inflammation is sustained by microglia activation to phagocytes that release pro-inflammatory cytokines. Research in the field of neuronal anti-inflammatory drug screening and technology is hampered by the difficulty in standardized cell culture and activation protocols. Also, it would be desirable to have guidelines for high-thorughput screening of compounds in loads of individual cells at hand alongside with screening image processing algorithms. The initiative for this new collaboration between two technical institutes aims to (i) optimize cell culture cell culture conditions of microglia cells isolated from mammalian brains in 2D bioreactors, (ii) to develop processes for quick controlled detachment of microglia cells to be harvested for high throughput microscopy, (iii) to develop a cell seeding technology to transfer microglia cells into multi-plate wells, (iv) to use long-term cell culture transillumination/epifluorescence microscopy to observe microglia activation cell changes in cell shape upon inflammatory cytokine stimulation over 12-36 hrs in individual cells in an automated imaging environment, (v) to perform second-harmonic generation microscopy of tubulin networks in activated microglia cells and (vi) to screen various promising new compounds for their ability to prevent microglia activation, e.g. cone shell toxins, green tea extracts, anti-inflammatory agents(minocycline). During the collaborative exchange project, key technologies will be developed to address these specific aims which will be pursued in subsequent grant applications on a larger scale. From the outcome of the project, we expect standardized protocols to investigate microglia pharmacology and pathophysiology on a biotechnological scale that might prove suitable for further industrial exploitation. Both partners are experts in single cell physiology, biophysics and biotechnology and will use latest state-of-the art imaging procedures with long-term cell culture microscopy.