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English
Views: 0 Author: Site Editor Publish Time: 2024-05-15 Origin: Site
In cell experiments, cells not adhering to the wall is a common and troubling problem. It often brings a lot of inconvenience to the experiment and affects the experiment and the accuracy of the results. When encountering a situation where cells are not adhering to the wall, we can start from the following aspects to investigate the reasons for the non-adherence and find corresponding solutions.
01 Reasons and solutions for cells not adhering to the wall
Trypsin overdigestion
microbial contamination
Mycoplasma infection or other microbial contamination can cause cell dysfunction and affect adhesion ability;
Solution: Isolate the culture and test for mycoplasma. If mycoplasma contamination is found, the culture should be discarded.
Medium pH too alkaline
If the pH value of the culture medium is too high, it may cause the cells to become unadapted and affect their adhesion;
Solution: Use sterile acetic acid solution, adjust the pH value or fill with sterile CO₂ to maintain a suitable pH environment.
cellular aging
Excessive cell passage times may cause cells to age and lose their adhesion;
Solution: Resuscitate new cells, use 20% serum to help cells recover and pay attention to the cell density and timing of subculture.
Improper starting concentration of seeded cells
If the cell inoculation amount is too low or too high, it may not be able to form an effective extracellular matrix and affect adhesion;
Solution: Change the composition of the culture medium and use a medium containing adhesion-promoting factors.
02 How to determine whether cells are contaminated
Look at the turbidity and color changes of the culture medium
Bacterial and fungal contamination often causes the medium to quickly become turbid and change color.
Observe under the microscope
The structure of bacteria and fungi can be observed with an intercellular microscope, helping to detect microbial contamination at an early stage.
Odor in culture medium
Certain microbial contaminations produce distinctive odors that are easily detectable, which can aid in the detection of contamination.
03 Precautions
During cell culture, aseptic techniques must be strictly followed to prevent cells from being contaminated. Use fresh and quality media and cell lines to reduce the possibility of contamination. Cell status also needs to be checked regularly, and if necessary, cells should be passaged or abnormal cells should be dealt with in a timely manner.
