Isolation and Analysis of Neural Crest Cell Subsets Using Fluorescence-Activated Cell Sorting and Cell Culture

Abstract

The ability to analyze the developmental behavior of defined cell populations is an important tool for understanding several questions concerning how cell commitment and differentiation are established during the maturation of the nervous system. The application of fluorescence-activated cell sorting and cell culture to the study of the mechanisms of differentiation of avian trunk neural crest cells is described. Results from these experiments show that this method is useful for the isolation and analysis of subsets of precursors defined on the basis of their ability to bind specific antibodies. In particular, findings indicate that cell surface differences expressed on young neural crest cells are correlated with the later development of particular differentiated phenotypes. These results are consistent with a combinatorial hypothesis in which the generation of phenotypic diversity in the neural crest results from the interaction of distinct subsets of progenitor cells (defined by the display of distinct arrays of cell surface receptor molecules) with particular ligands (e.g., growth factors and extracellular matrix molecules) In the embryonic environment. As a consequence of different sets of receptors and ligands combining on discrete subpopulations of neural crest cells, different pathways of differentiation would be elicited in particular subpopulations of cells. In this model, the action of cues in the embryonic environment is superimposed on existing developmental biases in particular subsets of neural crest cells.