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FAQs about Cell Freezing and Resuscitation Experiments

2024-01-29

1. Should DMSO be removed immediately when thawing cell freezing tubes for culture? 


Except for a few cells that are specifically stated to be sensitive to DMSO, most cell lines (including suspension cells) should be thawed and placed directly into a culture corner bottle containing 10-15 ml of fresh medium, and then replaced with fresh medium the next day to remove the DMSO, which can avoid most of the problems that cells cannot grow or adhere to after thawing. 


2. Can I use a different medium from the original culture conditions?


No. Each cell line has its own specific culture conditions. Each cell line has its own specific cell culture medium that it has adapted to. If we suddenly use a medium that is different from the original culture conditions provided, most of the cells will not be able to adapt to it immediately, resulting in the failure of the cells to survive. 


3、How should I thaw the frozen tubes? 


After removing the frozen tubes, they should be thawed immediately in a 37 °C water bath. Shake the tubes gently so that they thaw completely within 1 minute, and make sure that the water does not rise above the edge of the lid of the tubes, otherwise contamination is likely to occur. When thawing the tubes from the liquid nitrogen tank, it is necessary to pay attention to the safety of the frozen tubes to prevent them from bursting. 


Can I use a different type of serum from the original culture conditions?


No. Serum is a very important part of cell culture. Serum is an extremely important source of nutrients for cell culture, so the type and quality of serum will have a great impact on cell growth. Serums from different species differ in the amount of some substances or molecules or in their contents, and the wrong use of serum will often result in cell failure. 


5、What is FBS, FCS, CS, HS ? 


FBS (fetal bovine serum) and FCS (fetal calf serum) have the same meaning, both of them refer to fetal bovine serum, FCS is a wrong word, please do not use it again, CS (calf serum) refers to calf serum, HS (horseserum) refers to horse serum. 


6. What are the ingredients of cell freezing medium? 


The most commonly used freezing medium for animal cell cryopreservation is a homogeneous mixture of 5 - 10 % DMSO (dimethyl sulfoxide) and 90 - 95 % of the original fresh medium used for cell growth. Note: Since DMSO gives off a lot of heat when diluted, DMSO should not be added directly into the cell culture, but must be prepared before use.