2 replies to this topic

[NeuroADResearch]

Hi,

I'm trying to decide which cell line to use for Alzheimer's disease (I am transfecting through electroporation APPswe).

I want to investigate the effects of a chemical on Aß42, tau proteins, and reactive oxygen species. I know that APPswe does show both Aß42 and tau phosphorylation, and ROS. I am not sure if I should use a microglial cell culture to study the effects of the chemical on ROS, because the ROS are mainly studied with microglial cells.

What have you heard or know is the best cell line for this cell culture for AD, HEK293 or SH-SY5Y?


Also, what protocol would you recommend for the medium of that cell line? Serum-free/with serum?


Thanks! Any further advice would help!

[Zee]

Have you considered testing both lines? I always like it when data is replicated in 2 different kinds of cell lines in publications. Personally, I would start with HEK293 and then confirm the results in SH-SY5Y if it looks promising. This is because HEK293 is easy to obtain, grow, and manipulate for experiments.

In different labs, I've grown HEK293 in DMEM+10% FBS (heat-inactivated)+1%Pen/Strep and in DMEM/F12 50/50+10% FBS (heat-inactivated)+1%Pen/Strep. The cells grew faster in the DMEM/F12, but that could have been a stock difference.

Note: The origin of HEK293 is a topic of debate. It expresses many neuronal markers and has been hypothesized that it originates from a neuronal lineage rather than the kidney tissue from which it was isolated (Shaw,G., et al, 2002). While historically some labs have used it as a negative control and compared it to neuronal lines, it is not the best model. That being said, I think it makes a great tool for neurobiologists & researchers looking at AD.

[NeuroADResearch]

Yes! I didn't think of that, it won't hurt to try both.

Thanks!