Dear Noora!

I also work with neurons.

There are compounds - growth factors, for example, nerve growth factor, which are not toxic to normal neurons and induce differentiation in them (maturation, maintain survival).

Article Differentiation by nerve growth factor (NGF) involves mechan...

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Dear Alexandr,

Thanks for the answer! During neuronal maturation, I have added BDNF, GDNF, and CNTF in the medium.

Best, Noora

Dear Noora Räsänen

Adding inhibitors has been a traditional way to reduce NPC proliferation / glial proliferation. The traditional media also has high glucose concentrations that promote survival but isn't necessarily physiological. Another approach to this problem is to provide more physiological media such as found in vivo.

STEMCELL Technologies launched BrainPhys media in 2015, based on research from Cedric Bardy in the Fred Gage lab. BrainPhys is based on human CSF, and promotes activity and maturation.

Original BrainPhys paper:

https://www.pnas.org/content/112/20/E2725

What this means for primary-derived neurons

https://pubmed.ncbi.nlm.nih.gov/29199132/

What this means for PSC-derived Neurons

https://www.sciencedirect.com/science/article/pii/S1873506119301710

We hypothesize this is related to activity-dependent maturation mechanisms. Fire-together, Wire-together?

Thank you Jason,

I actually use additional glucose in the medium according to the original protocol. I am also familiar with BrainPhys, and we might even have it in the lab.

I have now realized that FUDR, that I've mainly used to inhibit proliferation, works with a delay. It takes two days for it to kill the proliferating cells, and now this has been coinciding with the plating day. I'm now trying to apply it at an earlier time point. I may have to accept the fact that most of my cells are resistant to differentiation.

Best, Noora