engleski

Neuroblastoma cells express glial features during differentiation

The neuroblastoma cell line SH-SY5Y is widely used as a neuronal cell model to study mechanisms of neurodegeneration, such as those in Parkinson's and Alzheimer's disease. In most protocols for SH-SY5Y cell line differentiation, one week all-trans retinoic acid (RA) treatment as a single differentiation factor is applied. The differentiation into "nearly pure" neuron-like cells is confirmed by analysis of neuronal markers expression and cell morphology. However, the presence of glial markers in these differentiated cells is being neglected in both classical and more current studies. As a consequence, biochemical data on coexisting lineages of glial and neuronal phenotypes in SH-SY5Y cell cultures is missing. Here we report our results on expression of glial fibrillary acidic protein (GFAP), pan-neurofilament proteins (NF) and the neuronal isoform of beta-tubulin in SH-SY5Y cells, as revealed by immunocytochemistry. In the group of undifferentiated cells nearly all of the cells stained concomitantly for GFAP and NF (93.1%) or GFAP and tubulin (95.9%). In these two subgroups only a small proportion of cells were exclusively GFAP- (5.8%), tubulin- (3.6%) or NF-positive (1.2%). On the other hand, in the group of differentiated cells treated with 10 nM RA there was a slight, but statistically significant increase in cells exclusively immunolabeled for NF (5.8%) and tubulin (9.4%). At the same time, the difference in the total number of GFAP-positive cells between the groups of undifferentiated and differentiated cells was not significant. These results implicate that during differentiation a significant number of SH-SY5Y neuroblastoma cells maintain some glial features, suggesting a slower and less stepwise process of their neuronal differentiation than previously thought.

beta-tubulin; cell culture; differentiation; glial fibrillary acidic protein; neuroblastoma; neurofilaments; SH-SY5Y

nije evidentirano