We have been following protocol which explains how to obtain cortical neuron from iPSCs. Despite having really promising results, we have mixed culture, so far. I am wondering if I can use mitotic inhibitors during this developmental process such as either using combination of FdUr, rand1, araC or using them separately?
Thanks advance
I never tried using such inhibitors, so I am not be able to guarantee the result. But if you would like to try, why not?
I couldn't be sure whether using inhibitors would be toxic to cell or not. But I am planing to use then I will share my observation. And hopefully somebody may give suggestion.
I have found AraC to be toxic to the differentiating neurons. I don't have a better solution for mixed cultures, but I think that starting the differentiation at higher density may improve the percent neurons.
Ara-C and FUDR are regularly used in primary neuronal culture to inhibit the proliferation of glial cells. They are generally not toxic to the developing neurons when applied at ~1-4 days after plating, or even sooner. I think you can have a go with them.
I haven't used those inhibitors before, but I've used cell cycle arrest drugs, DAPT and CDK5 inhibitor: PD0339221 (I think that's the right number). These were introduced at a few weeks post neural induction of the ips. Worked well with no adverse effects
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