Proper function of the nervous system depends on communication through neurons via axons and dendrites. The formation of these specialized cell processes first begins with neurite outgrowth. Measurement of neurite outgrowth is a convenient method to gauge neurotoxicity in screens for drug safety as well as a drug discovery tool for diseases of neurodegeneration.
Typical neurite outgrowth assays are standardized to a model system using a cell line that can reliably be induced to grow neurites in response to stimuli such as nerve growth factor. Cells are then exposed to compounds for a set period of time and neurite length is measured by immunofluorescence microscopy, often in high throughput using high content imaging platforms.
The BioFlux system for live cell imaging (Figure 1) consists of microfluidic channels (Figure 2) optimized for controlled and rapid reagent exchange, cell growth and high quality fluorescence microscopy. For neurite outgrowth assays, the flow can be utilized to rapidly perform staining protocols without disturbing the monolayer, as is often the case in microti- ter dishes. Neurite monolayers often do not adhere tightly to culture surfaces. A quick bolus of perfusion from a pipette can damage this adherence. The system also allows real-time observation of cell growth under controlled conditions. BioFlux can be used for axon guidance experiments under precisely controlled flow conditions.
Here we report the results of a model neurite outgrowth assay using rat PC12 cells exposed to nerve growth factor (NGF) and cisplatin using a high content analysis kit to score the neurite growth.