Investigation of cellular immune response against neurotoxins in neurospheroid-based cerebral tissue-on-a-chip platform integrated with blood-brain-barrier
Project no. 22187 · Status: Kapanmış · 2020
Summary
<p>The complex structure of the human brain, which regulates all functions in the body by controlling the central and peripheral nervous system, complicates the understanding of the disease mechanism and the development of therapeutic approaches on in vivo model organisms in preclinical studies. In this context, three-dimensional (3D) in vitro tissue culture models developed for physiological and Functional mimicry of the cerebral cortex, the largest neural integration region, are extremely important. The aim of this project is to investigate the neurotoxic effects of sedative agents at the cellular level on a new 3D cerebral organoid-based and blood-brain barrier (BBB) supported new design 3D-cerebral-on-chip platform consisting of induced pluripotent stem cell (iPSC)-derived neuronal cells. In the method to be followed in order to achieve the goal, firstly, neuron, microglia, mast, astrocyte and oligodendrocyte cell differentiation-characterization-isolation processes from iPSCs are performed. An extracellular matrix structure is formed with the brain tissue to be decellularized together with mast cells. Then, the microfluidic chip platform is fabricated, where endothelial cells and 3D cerebral tissue to support the BBB are cultured together. Optimization of dynamic culture conditions and engineering calculations are made. Next, the cellular effects of the neurotoxins to be tested on the cerebral-on-a-chip platform are examined by cell viability-apoptosis, immunofluorescence, western blot, qRT-PCR, ELISA, and glutamate assays. In parallel, at the Ludwig Maximilian University, Helmholtz Zentrum Munich, with the laboratory infrastructure and support of Dr. Ali ERTÜRK, tissue cleaning techniques are applied, and image optimization and analysis are made under 2-photon microscope of 3D cerebral tissues before and after neurotoxin application. In this way, the damages that might occur in the cells as a result of neurotoxicity in the 3D cerebral tissue, emulated in the microfluidic platform have been examined in depth with innovative/advanced imaging techniques.</p>