Urgent question! Do human cells survive at -20 degrees for couple of weeks or do we have to be store them at -80 or below. Any suggestion will be greatly appreciated?
Hello Sıla Köprülüoğlu
If you are going to culture these human cells in future (after a few weeks), then you may have to store them at -80 degree C or in liquid nitrogen but in a freezing medium containing 90% complete culture medium + 10% DMSO. You will also have to freeze them gradually at -1°C/minute using Mr.Frosty container. This container is designed to achieve a rate of cooling very close to -1°C/minute, the optimal rate for cell preservation.
Human cells will not survive at -20-degree C.
Best.
Human cells can survive at -20°C for a short period of time, but it is not recommended to store them at this temperature for an extended period of two weeks. At -20°C, the cells will start to freeze slowly, and ice crystal formation can cause damage to the cell membrane and DNA. Additionally, the metabolic activity of the cells will decrease significantly, which can affect their viability and functionality.
It is generally recommended to store human cells at a temperature of -80°C or lower to ensure their long-term viability and stability. At this temperature range, the cells will remain frozen and their metabolic activity will be minimal, which helps to preserve their integrity and function.
If you need to store cells for a longer period, it's better to use liquid nitrogen (-196°C) or a cryopreservation method that uses a cryoprotectant as suggested by Dr. Malcolm Nobre to protect the cells during freezing. Cryopreservation methods can help to prevent ice crystal formation and cell damage, allowing the cells to survive for longer periods of time
You probably should add some glycerol to your freezing solution.
Thank you so much for your kind answers, I will let you know how it goes!
The simple answer is NO! Even if they have been frozen with DMSO they will change (even morphologically). I know the very hard way !!! In my case, adherent cells began to grow only in suspension and lost adherence property. Also, you can have significant cell death and recover only a fraction and it will in essence be a "selection" from within the population.
If they are irreplaceable cells, and critical, you can grow them. BUT to use them you must (1). Have not much loss in viability so you can recover a good representation of the population. Otherwise you are selecting for survivors.(2). Have previous assays that you can confirm that the cell behaves phenotypically as expected. Like a drug response, response to growth factor etc. This is critical if you want to reproduce previous results you got with the cell line.
Even otherwise, if they are critical cells and recoverable, you should note all the steps you perform to recover them, and report that accordingly.
Good luck!
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