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Neurite Outgrowth & Network Dynamics


Key Features & Benefits

  • Physiologically relevant readout. Human iPSC-derived neurons cultured in defined growth conditions.
  • High-content analysis. Neurite length/area & branch points measured at the single-cell and population level.

  • R2G assay services are a starting point. Transition to more complex, bespoke assay services with the same service provider.

Assays Designed with Your Goals in Mind

Assessing the impacts of therapeutics targeting neurological disorders on neurite outgrowth or neuronal network integrity is essential to the success of any neurobiology-focused, drug-discovery/development campaign. Additionally, identifying potential neurotoxic effects of therapeutics targeting other non-neuronal disease classes is beneficial in flagging and addressing undesirable, off-target effects that could jeopardize clinical advancement and, ultimately, patient safety.

The Challenge

As iPSC-derived neuronal cells grow and proliferate in vitro, the neurites tend to overlap extensively, making accurate measurements of neurite length, branch points, etc. difficult or impossible. As a result, accurately measuring the impact of therapeutics upon neurite outgrowth requires extensive optimization of cell density to achieve minimal overlap in neurites originating from adjacent cells while maintaining a sufficient density to ensure cell health.

While these assays can be relatively straightforward to establish using immortalized cell lines such as PC-12 cells, those models may not be as physiologically relevant compared with human iPSC-derived neuron models.

Regarding detection, despite broad availability of robust detection antibodies for neuronal cell markers, a standard, antibody-based detection approach typically requires extensive wash steps that could mechanically disrupt the neuronal network.

Our Solution

To address these challenges, PhenoVista has developed the Ready-2-Go (R2G) Neurite Outgrowth Assay Service and the R2G Neurite Network Dynamics Assay Service, enabling robust, accurate quantification of neurite outgrowth or network formation, respectively, using human iPSC-derived neurons and a homogeneous assay workflow with dye-based detection. We have optimized cell-seeding densities to ensure accurate neurite measurements, with the flexibility to assess network formation.

How It Works


Cells are seeded into 384-well plates. For neurite-outgrowth assessment, cells are incubated in the presence of drug for 4 and 24 hours, when they are fixed and stained for imaging and analysis. For assessing neurite network, cells are treated with drugs on day 6 for 24 and 72 hours, and they are fixed and stained on days 7 and 9 for imaging and analysis. In the images below, human iPSC-derived glutaminergic neurons were treated with reference compounds, and neurites were measured at 24 hrs. Nuclei were stained with Hoechst (blue) and cell bodies and neurites with CellMask (green).


Assay Service Details

  Ready-2-Go Neurite Outgrowth & Network Dynamics^ Bespoke Assay Services  
Cell Type iCell Glutaneurons (Fujifilm Cellular Dynamics, Inc.) If you would like to expand the service offering beyond R2G shown on the left, please contact us.  
Markers Hoechst (nuclei), CellMask (cell body/projections)  
Dosing 6 doses of your test article  
Positive Control* Staurosporine  
Negative Control* Vincristine  
Time Points Neurite Outgrowth^: 4, 24 hrs
Network Dynamics^: 7, 9 days
Assay Readouts Total cell count, neurite length/area, # branch points  

*Vehicle and untreated controls also included.
^Neurite Outgrowth and Network Dynamics are distinct service offerings and can be purchased separately.