Differentiation of the human NT2 cells into neurons and glia

A method underlying a strategy for differentiation of the NT2 human teratocarcinoma cell line into neuronal and glial cells is described. The aim of this work is to provide a human model to study the relationships between neurons and glia in vitro during developmental or degenerative events. NT2 cells are seeded on polylysine precoated plastic or glass and differentiated by all-trans retinoic acid; persistent undifferentiated cells are eliminated by cytosine--D-arabinofuranoside; then cell cultures are maintained during four weeks until the appearance of glutamatergic receptors. Along the differentiation procedure, we have followed the expression of neuronal and glial phenotypes as well as the excitotoxic response to N-methyl-D-aspartate treatment taken as an indication of neuronal maturation. The procedure described leads to the development of a mixed population of neurons and glia sensitive to glutamate exposure.