Neurite Outgrowth Assay Services for Stroke Drug Discovery & Development

Neurite outgrowth is the extension of the developing neuronal cell body into numerous dendrites and axons and is regulated by complex intracellular signaling events. Nerve growth factors or neurotrophins are one of a family of stimulators that regulate neurite growth. Dynamic neurite outgrowth during development leads to the formation of complex neuronal structures that form a functional nervous system and brain. Regulation of the neurite outgrowth process has been implicated in a variety of central nervous system disorders, including neurodegenerative disorders and stroke. Understanding the biology of neurite outgrowth can reveal the underlying mechanisms of these diseases.

Fig. 1. Effect of rosuvastatin on neurite outgrowth in cultured cortical neurons. (He et al., 2018)

Custom Neurite Outgrowth Assay Services

Ace Therapeutics is a leading CRO in the field of stroke research, offering comprehensive neurite outgrowth assay services tailored specifically for stroke drug discovery. Our neurite outgrowth assay studies the potential effects of stroke drugs on neuronal activity and development in vitro, providing valuable information on the potential of stroke drugs to enhance neurogenesis.

Using in vitro models of ischemic stroke based on induced pluripotent stem cells (iPSC) and neuronal cultures, we can study the segmentation and quantification of neuronal processes to assess neurite growth. These neuronal processes can be imaged using fluorescence microscopy and quantified by manual tracing and counting at low fluence.

Our high-content imaging team of neurons can accurately characterize and measure changes in neuronal networks, including:

• Neurite quantification
• Number of neurite nodes
• Neuronal bodies quantification
• Neurite length (maximum & average length)
• Determination of overall or specific toxic responses

Workflow of Neurite Outgrowth Assay

• Culturing neuronal cells: Neuronal cultures are established in 96 or 384 well microtiter plates to form a network of neurites.
• Treatment with compounds: Neuronal cultures are incubated with stroke drugs to be tested.
• Immunostaining and imaging: We use fluorescent antibody pairs to stain neuronal microtubules, allowing visualization of neuronal synapses. A high-content imaging system captures images accurately and in detail.
• Automated analysis: We use cellular imaging analysis software to quantitatively analyze neuronal cell images to characterize multiple parameters, including the number of processes per cell, length of synapse growth, branching, and cell number.
• Results Readings: If the compounds show the ability to shrink neurite growth, then this would be an indicator of neurotoxicity. On the other hand, if the compounds show the ability to produce or elongate these protrusions, then this would be an indicator of neurogenesis.

Ace Therapeutics's neurite outgrowth assay studies neuronal development and neuronal degeneration in vitro, making it possible to identify effective stroke drugs. Our laboratory has a state-of-the-art automated imaging system used in conjunction with analytical software to assess the effects of stroke drug compounds on cell viability and neurite outgrowth dynamics. If you are interested in our services, please do not hesitate to contact us!

1. He, W., et al. (2018). Rosuvastatin improves neurite outgrowth of cortical neurons against oxygen-glucose deprivation via Notch1-mediated mitochondrial biogenesis and functional improvement. Frontiers in Cellular Neuroscience, 12, 6.