Hi everyone,
i might have a dumb question... my fish cells keep detaching and dying after a few days of culture so no i am trying to find a solution to this problem.
They grow at 19°C without CO2. I only have experience with mammalian cells. I bought a new incubator, which has no extra water tank. Now my question is: Do i need to place a tray or something in the incubator, even if the cells grow at low temperature? Might this be the reason they keep dying?
Hello Anika Dreier
Usually, a tray filled with sterile water is placed in the incubator to generate a humid atmosphere inside. This atmosphere will help to maintain growth-friendly conditions for the cultured cells. If the tray with water is not present in the incubator, then without much humidity over time, the osmotic balance in the cell may be lost and the salt concentration of the media in which your cells are growing may increase, thereby having detrimental effect on the cells.
So, even at a temperature of 19-20 degree C, I feel you should keep a tray of sterile water in the incubator containing some additives (disinfectants) to prevent contamination.
Best.
For salamander cells growing at 18C, I use a 150 mm outer dish with 3x35mm culture dishes around a 35 mm water only dish. All with lids on.
Ellen A G Chernoff Thanks for the tip! Could you please tell me, why you keep the lids on? I now put 4 Petri dishes with 25ml each on the bottom of the incubator but without lids.. would that be ok? Or could it be "too much"?
Anika Dreier. I use the lids to prevent spills when moving the whole thing.
The try used to saturate the air with water should be of the same osmotic strength as the medium the cells are in. If this is not done there will be a slow transfer of water from the osmotically weaker to the stronger solution. Depending on how frequently you change the medium this may or may not be a problem. The pH of the media may also increase due to CO2 loss therefore the atmosphere above the cells should have the appropriate CO2 concentration to circumvent this effect.
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