Neurite Integrity Assay with Brain Extracts

Human iNs of glutamatergic cortical fate were differentiated for 21 days before the assay. Cells were imaged every 2 or 4 hours using an IncuCyte system (Sartorius) and treated with brain extract or cell-derived lysates. Neurite integrity was quantified using the IncuCyte NeuroTrack Software Module (Sartorius). Total neurite length per well were normalized to the values from the first 6 hours of imaging. Additional details relating to this assay were previously published [6 (link)].

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Hsieh Y.C., Negri J., He A., Pearse RV I.I., Liu L., Duong D.M., Chibnik L.B., Bennett D.A., Seyfried N.T, & Young-Pearse T.L. (2022). Elevated ganglioside GM2 activator (GM2A) in human brain tissue reduces neurite integrity and spontaneous neuronal activity. Molecular Neurodegeneration, 17, 61.

Publication 2022

IncuCyte system IncuCyte NeuroTrack Software Module

Assay Brain Cell Cortical Human Neurite

Corresponding Organization : Harvard Stem Cell Institute

Other organizations : Brigham and Women's Hospital, Emory University, Massachusetts General Hospital, Rush University Medical Center

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Variable analysis

independent variables

Brain extract
Cell-derived lysates

dependent variables

Neurite integrity
Total neurite length per well

control variables

Human iNs of glutamatergic cortical fate cells
Differentiation time of 21 days
Imaging frequency of every 2 or 4 hours using IncuCyte system
Neurite quantification using IncuCyte NeuroTrack Software Module

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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