NickSarbiscuit

Thanks for the advice!

I'll pass it onto the lab and we'll do some more CO2 checking. The issue with the incubator is that it's CO2 monitor seems to be off, it registers as the correct CO2, but the centre tech measured with his own monitor and came up with a different reading. We'll do a third test to make sure it's not the problem. the incubator is meant to be at about 6% CO2, how low do you think it would have to be until the media fails to change colour?

In terms of the cells. They are not immortal and don't replicate a much if at all (past a certain stage). The Glia have been passaged twice, and won't be passaged again (although it is possible, I wouldn't do more than 3/4 times as past that they won't grow at all). The Neurons on the other hand, if they have lost their pluripotency and are truly turning into neurons, passaging tends to tear them apart so can't be done much at all!

Considering the work we are doing (hiPSC derived neurons from patients) the cells are generally difficult to work with and extremely valuable so can't really do any side-testing. May be able to do something with a different sample.

Just to throw in my 2 cents. In a neuro lab we spin at 90g for glia, and 160g for neurons in 15ml falcon tubes for 3 minutes.

Considering neurons are pretty small I doubt you'd need to go much faster.