Hi,

For SH-SY5Y protocol, you can go through our publication

" Differentiation Induces Dramatic Changes in miRNA Profile, Where Loss of Dicer Diverts Differentiating SH-SY5Y Cells Toward Senescence"

this works good and used by other researchers as well

Hi Terezie Páníková

I have been working on differentiated SHSY-5Y cells. I would suggest you to use Retinoic acid (10uM) in 1% heat inactivated medium in F12 medium (or the media which your cells are conditioned to). Change the media every 2 days and the cells would be differentiated in 7-8 days. You can confirm the same by immunoblotting of Tyrosine Hydroxylase.

Thanks

Hope it works for you.

For the basics and general cell culture, as well as preparation for efficient differentiation, I recommend:

Feles, S.; Overath, C.; Reichardt, S.; Diegeler, S.; Schmitz, C.; Kronenberg, J.; Baumstark-Khan, C.; Hemmersbach, R.; Hellweg, C.E.; Liemersdorf, C. Streamlining Culture Conditions for the Neuroblastoma Cell Line SH-SY5Y: A Prerequisite for Functional Studies. Methods Protoc. 2022, 5, 58. https://doi.org/10.3390/mps5040058

For differentiation:

Kovalevich, J.; Langford, D. Considerations for the use of SH-SY5Y neuroblastoma cells in neurobiology. In Neuronal Cell Culture; Springer: New York, NY, USA, 2013; pp. 9–21.

Shipley, M.M.; Mangold, C.A.; Szpara, M.L. Differentiation of the SH-SY5Y Human Neuroblastoma Cell Line. J. Vis. Exp. 2016, 108, 56193.

Constantinescu, R.; Constantinescu, A.T.; Reichmann, H.; Janetzky, B. Neuronal differentiation and long-term culture of the human neuroblastoma line SH-SY5Y. In Neuropsychiatric Disorders an Integrative Approach; Springer: New York, NY, USA, 2007; pp. 17–28.