Hi all,
I am trying to differentiate SHSY5Y human neuroblastoma cells into neurons with the retinoic acid protocol. I add 10uM retinoic acid into 3% FBS containing DMEM for three days, and therafter, maintain them in 1% FBS-containing DMEM (also with 10uM RA).
I have done this for 13 days already, and while I do see some differentiation happening, many of the cells are still undifferentiated. I have read other papers which were able to see neuron like network projection neurons in about 4-5 days.
Should I increase the concentration of retinoic acid used to increase the rate of differentiation? I have Attached a picture of my D13 differentiated vs untreated control cells for reference.
Thanks and Regards,
Mathangi
Hi Mathangi Palanivel
The SHSY5Y neurons are differentiating according to the attached images, however still you can increase the conc. to 20uM and check for the neurite growth. Once you observe neurite projections, stain with Tyrosine Hydroxylase to check the its expression levels (marker for differentiated dopaminergic neurons).
Hope it helps.
Thanks
Samir
Hi we use a 5 day treatment with RA and then a 6 day treatment with BDNF during differentiation. This results in cells that look a lot more neuronal.
Tobias Karlsson oh ok thanks you! May I check what were the respective concentrations of RA and BDNF that you use? Also, when changing to BDNF treatment, do you maintain the RA in the medium?
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