Hello all
I'm working with SHSY5Y cells and would like to differentiate them to neuronal cells. I used differentiate media with Retinoic Acid (RA) for this. After finishing the incubation period, I checked the cells under microscope to see the morphology changes after differentiate time. By morphology, I could see the difference between Un-differentiate and differentiate cells; presence of long, branching neurites, more elongated and neurite-bearing morphology compared to undifferentiated cells.
My question is beside morphology changes, how I can make sure my un-differentiate SHSY5Y cells become differentiate to neuronal cell and they are mature? For example check functional change or any other changes.
Thank you
Hi Elham Amini. E.A
Apart from the morphological differences observed such as the neurite growth. Cells fully differentiated can also be stained with markers such as growth-associated protein (GAP-43), neuronal nuclei (NeuN), synaptophysin (SYN), synaptic vesicle protein II (SV2), neuron specific enolase (NSE) and microtubule associated protein (MAP).
Thanks,
Dear Samir Ranjan Panda,
Thanks for your reply. Apart from staining, can I use these antibody such as neuronal nuclei (NeuN) to do western blot as well?
Hi Elham
Yes, you can confirm the same markers through western blot.
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