Application Note

Visualizing Neurite Outgrowth by Lentiviral Transduction of Fluorescent Proteins Into Human iPSC-Derived Excitatory Neurons

Label nearly 100% of hiPSC-derived excitatory neurons with GFP reporter lentivirus for live-cell imaging of neurite outgrowth, excitotoxicity assessment, and long-term calcium oscillation monitoring.

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Inside this application note

Review lentiviral transduction conditions, optimal MOI data, and live-cell imaging results for neurite outgrowth visualization, glutamate excitotoxicity, and GCaMP6f-based calcium oscillation monitoring in hiPSC-derived excitatory neurons.

GFP expression detected in the majority of cells by day 8 post-thaw using a SYN1 promoter-driven lentiviral vector, with MOI 2.5–10 sufficient for clear visualization of thin neurites without significant toxicity
Severe neurite degeneration observed at 10 μM glutamate by day 21 post-thaw, with neurite beading identified as a potential sensitive early endpoint for neurotoxicity prediction prior to cell death
Spontaneous calcium oscillations detected from day 21 post-thaw using GCaMP6f, with the most stable signals at approximately 1 peak per minute obtained at day 29 post-thaw at MOI 30

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