Hello Research Gate,
I am planning on using a quite specific cell line which has been immortalized with the temperature sensitive SV40 T antigen. Cells immortalized this way are widely reported to grow at the "permissive" temperature of 33C and to stop dividing and die/differentiate (depending on the cell type) at the "non-permissive" range of 37-39C. My question is, has anyone used cells transformed in this manner, and if so, do you know how they do between 33C and 37C? Have you ever tried growing cells like this at, say, 35C or 36C?
The issue is I want to attempt a co-culture experiment with cells that are not transformed, so if possible I would like to find a temperature range that is amenable to both cell types. Thanks in advance.
I use the endothelial cells line obtained from SUVECs using SV40 T-antigen. In 36-37C they grow very fast. Much faster than primary cells. Morever they seem to be invulnerable for small deficiency of the medium components. Bui I've never tried to culture them in 33-35C.
Hi Stephan.
SV40 T is seldom used these days for obvious reason.
I can only suggest on the matter of co-culture system of temperature-sensitive cells (33C culture, mild hypothermic temp) and wild-type of normothermic temp cells. We have experienced cell culture at varying temperatures in both different incubators and a small sized-temperature controlled device having varying temperature controlled surfaces.
What we found was very active cell proliferation and hypoxia at normothermia, deteriorating rapidly. In contrast, cell proliferation is so slow, but with high cell viability, lasting for much longer term at 30~34C hypothermic condition. They were differentiating ES cells and neuroblastoma cells.
Thus, if you have lots of time and could bear the speed of doubling or differentiation or cell activation upon your trigger, you may choose the lower temperature in the middle as you put. The effects of temperature on metabolism, leading gene expression, is enormous beyond that we think.
Best wishes.
Hy,
I worked with temperature sensitive immortalised neuronal progenitor cells from the rat midbrain (CSM14.1-cells).
They proliferated also at 37°C (not only at 33°C) , but really only differentiated into a neuronal fate at 39°C when the serum was reduced from 10% to 1%. When we cultured them with 10% serum they continued to proliferate.
So, you have also to consider the serum content of your media and not only the temperature.
Sincerely Yours/Beste Grüße
Stefan (with an f)
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