Automatic Quantification Of Cell Outgrowth From Neurospheres

In the development of new regenerative medicine therapies for the treatment of central nervous system and spinal cord injuries, the identification of factors that inhibit or promote cell outgrowth in neurite outgrowth assays is fundamental, and the neurotrophic activity is commonly assessed based on the neurite/cell outgrowth. Neurites are projections from the cell body or the initial neurosphere and typically present low-contrast to background in phase contrast images. The extent of neurites is usually measured in a manual way and fluorescence images are the most used, generally requiring imunofluorescent staining.We present a novel automatic approach for the quantification of cell outgrowth from neurospheres, based on phase contrast and fluorescence images acquired from samples merely processed for DNA staining.Our approach detects the neurite/cell outgrowth, and its measures are in high agreement with the ones obtained manually. Furthermore, the image analysis time was reduced in more than 95% allowing the increase of the amount of data to be analyzed.