I am treating 1-week cultured SH-Sy5Y with 80 nM TPA for one week, replacing the media with the TPA twice a week according to (Presgraves, 2004). Instead of growing neurites, I notice that the cells actually become more spherical versus elongated and neuronal-like. Also, treatment with TPA is supposed to dramatically slow down cell growth. However, the cells appear to be growing at a faster rate than non-differentiated ones. Any ideas as to why this could be happening? I have tried differentiation with retinoic acid, but have chosen to differentiate with TPA instead.
Dear Brigitte, try to treat the cells first with retinoic acid alone for 3 days and then add TPA
I've only used all-trans retinoic acid to differentiate SY5Y cells, but in my experience it takes 7-10 days to see real changes in growth rate and neurite extension.
Also, be very careful with retinoic acid as it is highly teratogenic.
I have tried treatment with RA for 3 days, change media, then add TPA for 3 days as suggested by Arianna Bellucci. However, when I initially tried this when I first started using this cell line (low passage), this produced little morphological change and the cell growth only slowed down when TPA was added. Which is why I chose to move ahead with just TPA. Arianna, how long do you treat with TPA?
I have no experience with TPA or MPA. But we can differantiate the SH-SY%Y cells with retinoic acid.
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