I was wondering what effect does the variation in incubation temperature have on the growing cells, specifically neuronal cells? I read that mammalian cells should be incubated at 37 deg, but the incubator I will use can only be set to 35 deg due to some constraints. Will a 2 deg temperature difference matter for the cells?
Thanks!
From a metabolic point it would be the equivalent of hypothermia which won't reflect how these cells normally behave under test conditions
No, 35 is too low for optimal growth of cells, please try to fix your incubator.
a lot depends on the type of cells and the readouts you want to follow. growth may be slightly slowed, but synaptic responses may be affected. please note that mouse cells tend to be more resilient to hypothermia than human cells, and cell lines are far more resilient that primary cultures...
Thank you. I will be working with rat neuronal cell lines, and recording their response (action potentials) to external stimuli. Will 2 deg lower still affect my experiments? Our incubators are being used at 35 deg for another set of experiments..
Then I would suggest you wait until you can run at 37 degrees
PLEAD for another incubator or run your experiment in another lab where the incubator is set at the correct temp
There are several papers documenting the effect temperature has on action potentials
here's one I just found
Journal of Physiology (1995), 488.2, pp.407-418
Temperature effects on membrane potential and input resistance in rat hypothalamic neurones
JohnD. Griffin and Jack A. Boulant
This depends in what you are modelling with these neurons. The peripheral terminals of nerve fibres in the limbs may well be below 35o, perhaps as low as 30 or 32 in skin. If you want to model neurons of the CNS it is closer to 37oC. If the incubator temperature oscillates at all, the nerve cells will not fare well much above 37 so 35 might be a safer set point.
I wouldn't for a single moment trust the temperature setting of your incubator without first measuring your incubation solutions using a reliable thermometer independently. Aim for 36-37.5 degrees. If it's outside that range, adjust the setting. If you can't reach that range reliably or the temperature fluctuates outside the range, get a new incubator. There's no point wasting your preparations and time for a piece of kit that's not fit for your purpose.
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