Differentiation of SHSY5Y - NGF - 7S or 2.5S? (May/05/2009 )

Which should I use, 7S or 2.5S NGF? Or are they interchangeable? I am trying to promote neurite outgrowth.....

cmel on May 5 2009, 10:23 PM said:

ATCC doesn't recommend any NGF for this cell line. That said, I use NGF-B from Sigma-Aldrich for my primary neurons at a concentration of 30 ng/mL to promote neurite growth. I use the rat derived (N2513) but they also have human NGF-B (N1408). I'm not familiar with the nomenclature you're using (7S and 2.5S) but I hope this may be of some help.

gfischer on May 6 2009, 11:38 AM said:

Thanks for your reply,

I've got several papers which have done the experiment I'm after using this cell line but they just say 'NGF' was used at 10 ng/mL. I'm not really sure why you say that the ATCC doesn't recommend NGF for this cell line...

but, more to the point ..... you can get 7S NGF which is comprised of 5 subunits, or you can get NGFbeta (one of these subunits) or 2.5S NGF, which is an analogue of NGFbeta. I'm relatively new to this kind of work, so I'm not sure which is the appropriate growth factor to order! My suspicions are that NGFbeta or 2.5S NGF are the right way to go, but I'd appreciate some advice

Thanks again

ATCC probably doesn't say anything about NGF because its not required for growth of the cells, which is the main concern of the instructions they give. If none of the literature gives a specific type, then it's possible that it doesn't matter. At any rate, in the absence of further guidance, your best bet may be to just try each one and see what happens. Like I said, all I can tell you is that NGF-B works great for primary adult neurons. 10 ng/mL is a good starting point, but you may have to tweak the concentration for best results. Use the lowest concentration you can, because oddly enough, NGF becomes neurotoxic at higher concentrations.