ABSTRACT
PURPOSE: To determine the most effective method of dissociating neural stem and progenitor cells into a single-cell suspension. MATERIALS/METHODS: Induced pluripotent stem cells were differentiated toward the neural fate for 4 weeks before clusters were subjected to enzymatic (Accutase, trypsin, TrypLE, dispase, or DNase I) or mechanical (trituration with pipettes of varying size) or combined dissociation. Images of cells were analyzed for cluster size using ImageJ. RESULTS: Cells treated with the enzymes Accutase, TrypLE, or trypsin/EDTA, these enzymes followed by trituration, or a combination one of these enzymes followed by incubation with another enzyme, including DNase I, were more likely to be dissociated into a single-cell suspension. CONCLUSIONS: Cells treated with enzymes or combinations of methods were more likely to be dissociated into a single-cell suspension.
