Development of an in vitro model of neuronal differentiation and nerve-target interaction

Abstract

My objective was to develop a nerve source for use in a 3-dimensional in vitro model to study neurite extension and target innervation. A hybrid neuroblastoma-DRG cell line (ND-C) with properties of neuronal precursors was examined for its ability to effectively differentiate into neurons, produce extensive neurites, and interact with target cells. Neuronal differentiation of ND-Cs was optimized through a combinatorial approach that involved culturing the cells in the presence of various types of culture media, ECM, and soluble factors in a dose dependent fashion. The proportion of neurons and the length of neurites generated following culture in the differentiation promoting conditions were assessed. Immunostaining and RT-PCR analysis of differentiated ND-Cs revealed the expression of markers specific for terminally differentiated neurons. ND-Cs were able to extend neurties within a 3D collagen matrix. In co-culture, neurites from ND-Cs formed contacts with muscle and epithelial targets, indicating the plasticity of the cell line.