I've been trying to grow my HeLa cells from the cell stock for 3 weeks but it isn't growing at all..!
When I grew HeLa before, it was growing so fast that I had to subculture it every other day.. but now it dies and never grow..:(
What is the problem..?
Bacterial contamination, selection agent mix-ups, incorrect serum medium composition, etc. Various technical troubles are feared.
On the other hand, I will write about an example that is difficult to notice.
There are uncoated culture dishes for floating cells.
Did you use a culture dish for adherent cells?
Are you performing flask culture?
Depending on the cap shape of the flask, CO2 gas exchange may not occur easily.
If CO2 gas exchange does not occur and PH control does not work, the PH of the culture medium becomes alkaline and the cells die.
Is the temperature in the CO2 incubator properly controlled?
If the temperature sensor or heater is malfunctioning and the temperature remains above 38°C, strong stress on the cells will occur.
These are just a few things that come to mind.
Hello Hannah Hwang
I agree with Narumi Uno . Adding to the above answer.
Earlier HeLa cells used to grow well, and now they don’t. This means you need to re-look at the culture conditions provided to the cells. It is essential that the culture environment replicates, as best as possible. Culture conditions that you need to look into, and which can be controlled include temperature, relative humidity, and CO2 levels as well as factors associated with the media, such as nutrient composition (change of company manufacturing FBS), pH, osmolality and the volume and frequency of replenishment. These variables can fluctuate over time so they should be monitored.
Also,
Was the cell stock from a different lot not frozen following the proper freezing protocol?
Was the storage temperature maintained below -130 degree C (liquid nitrogen)?
Is the LN2 level in the storage tank carefully monitored in order to avoid the tank running empty.
These are important points which you need to look into in such a case.
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