Tracking the movement of individual avian neural crest cells in vitro

The origin, migratory pathways and adult derivatives of neural crest cells (NCCs) are well known. However, less is known about how these cells migrate. In this study, in a laboratory based in a low-resource setting, a hanging drop culture assay was utilised to study the movement of individual avian trunk neural crest cells. Mode of migration by means of lamellipodia and filopodia was studied in live cell cultures with a laser scanning confocal microscope and Airyscan module. Both distance migrated and speed of migration were calculated. NCCs migrated in a chain soon after emerging from the explanted neural tube, but were more dispersed and had random movements when they reached the periphery of the culture. While the distances travelled by these NCCs were less and the cells were slower on gelatine than on other extracellular matrices reported in the literature, the assay afforded detailed observation of actin filament distribution and cytoplasmic protrusions. The study has provided unique evidence of individual NCC movements in vitro, in a simple hanging drop assay optimized for the study of NCCs. The assay could be used for further analysis of the behaviour of NCCs on different extracellular matrices or with targeted action.