A novel green fluorescent SH-SY5Y cell line has been developed through stable transfection with tubulin tagged Evrogen TagGFP2. This cell line expresses green fluorescent tubulin marking the cell cytoskeleton and it can be easily differentiated using standardized protocols to promote the neurite outgrowth.Cells were then grown in the presence of G418. Single cells with strong green fluorescence were selected by flow cytometry, and allowed to expand. These cells constitutively express the TagGFP2-tubulin fusion protein.Assay Details: The growth of neurites in human neurons is a critical event in neuronal development, formation and remodeling of synapses, response to injury, and regeneration. Many neuroscience projects are focused on identifying new drugs that affect this event. The discovery of new compounds that can positively affect neuritogenesis would be very important for developing new therapeutics against both neurodegenerative diseases and injury. Measurement of neurite outgrowth using an automated image-based assay can be of use in the research, screening and validation phases of the drug discovery process. Moreover, a neurite outgrowth assay can also be used for testing toxic neuropathies. Recombinant SH-SY5Y cells stably expressing human tubulin TagGFP2 are intended to be use as in vitro model for neuronal differentiation studies, designed to assay for compounds that positively affect neuritogenesis quantifying the number and length of neurites.
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