PHENOTYPIC MORPHOLOGY AND THE EXPRESSION OF CYTOSKELETAL MARKERS DURING LONG-TERM DIFFERENTIATION OF HUMAN SH-SY5Y NEUROBLASTOMA-CELLS

A simple method for differentiating human-derived SH-SY5Y neuroblastom a cells to provide a stable, mature, neuronal morphology is described. SH-SY5Y cells can be induced to differentiate terminally with retinoi c acid in medium with low levels of serum. The morphological different iation of the parental cell line SK-N-SH was compared with that seen i n the SH-SY5Y cells. Changes in the cytoskeleton of SH-SY5Y cells indi cated that differentiation proceeds continuously over the 1-month peri od studied after initiating differentiation. Immunoblot analysis demon strated increased expression of the high molecular weight neurofilamen t polypeptide NF-H, the microtubule-associated protein tau, and the sy naptic vesicle-associated protein synapsin I, indicating that an incre asingly mature, neuronal phenotype was being expressed. The cultures w ere not dependent on retinoic acid for continued survival. SH-SY5Y cul tures differentiated over extended periods should provide a good in vi tro model for studying the neurotoxic potential of compounds and mecha nisms of toxicity, particularly in longer-term or multiple exposure st udies, for example on cytoskeletal function, where acute toxicity is n ot the aspect of interest. Copyright (C) 1996 Elsevier Science Ltd.