In vitro differentiation of neuron-like cells.

Abstract The pluripotent embryonal carcinoma, PCC4uva, differentiates into multiple cell types under standard culture conditions. Neuron-like cells first appear at 8 days of culture, and by Day 15 they represent approximately 3% of the cell population. We promoted neural differentiation by treatment with retinoic acid and dibutyryl cyclic adenosine monophosphate. Under these optimized culture conditions, neuron-like cells appear on the third day of culture, and by the sixth day, neuron-like cells represent 40% of the total cell population. After 10 days, the neuron-like cells represent approximately 60–80% of total cells. At this time the cells form large clusters of cell bodies that are interconnected by neurite fascicles. The cells express the neuron-specific proteins; Map2, Tau, neurofilaments, and the type III β-tubulin isotype. We have recently used this developing system to investigate the effects of laminin substrates on neural differentiation.