Contamination types, control and prevention of cell culture
Contamination types, control and prevention of cell culture
# Cell Culture
Contamination types, control and prevention of cell culture
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Cell contamination types and control
1. Bacterial contamination
Bacterial contamination is the most common type and is usually caused by improper operation. The bacteria appear spherical or rod-shaped under the microscope, which can cause the culture solution to turn yellow and cloudy, with an odor. The rapid reproduction of bacteria will rapidly consume nutrients in the medium and affect the growth of cells.
Treatment method
For bacterial contamination, it is possible to try to add antibiotics to the medium, however, it is important to note that the use of antibiotics may also have an adverse effect on the cells themselves, resulting in poorer cell status. Therefore, the most fundamental solution is still to strictly implement aseptic operation specifications, regularly clean and disinfect culture facilities, and focus on prevention.
2. Fungal contamination
Fungal contamination is difficult to detect early because it may not affect the color and clarity of the culture solution initially. Filamentous or coral-like structures may be seen under the microscope. Long-term fungal contamination can affect the normal growth of cells and even lead to cell death.
Treatment method
Once fungal contamination is found, it is recommended that the contaminated cells be discarded immediately and the entire culture environment be thoroughly disinfected and sterilized. This includes all potentially contaminated objects such as CO2 incubators, utensils and culture fluids.
3. Mycoplasma contamination
Mycoplasma is so small that it is difficult to see its morphological structure under conventional optical microscopy, and it cannot be removed by bactericidal filtration. They attach to the cell surface, causing the cell to deteriorate and grow slowly. Mycoplasma infection is difficult to detect, but can seriously affect the results of cell experiments.
Treatment method
For precious cells, treatment with mycoplasma removal media may be considered. This special component of the medium can effectively remove mycoplasma and reduce the loss caused by pollution.
4. Mold contamination
Mold pollution is manifested as flocculent impurities in the culture medium, and fine filamentous clumps of floating matter can be seen under the mirror, similar to catkins in the wind, which can not be detected without careful look.
Treatment method
Decisively discard the contaminated cells, and thoroughly disinfect the culture environment, scrub the CO2 incubator with alcohol and neogeramine, and add copper sulfate to the water tray to prevent re-contamination.
2. Cell pollution prevention
01
Aseptic technique
All cell culture-related operations should be carried out in a laminar flow cabinet to reduce the risk of contamination by airborne microorganisms. Work surfaces should be disinfected with UV light prior to operation, while ensuring that all reagents and materials used are sterile.
02
Media and serum quality
Using high-purity, tested media and fetal bovine serum, the quality of these products directly affects the success or failure of cell culture.
03
Maintain laboratory hygiene regularly
According to the frequency of use, regularly clean the laboratory and equipment, including ultra-clean tables, incubators, etc., to reduce the risk of pollution caused by the environment.
04
Physical and chemical pollution control
Control physical factors such as temperature and light in the laboratory, as well as the storage and use of chemical reagents to avoid adverse effects on cells.
05
Mycoplasma pollution control
Due to the small size of mycoplasma, it is difficult to observe and remove by conventional methods, so special mycoplasma removal reagents are needed for prevention and treatment.
06
Cross contamination prevention
When operating multiple cell lines, appliances and reagents should be strictly distinguished to avoid cross-contamination between cells.
Standard Operating Procedures (Sops) development: Establish standard operating procedures (Sops) to ensure that all operators can comply with regulations and reduce the risk of contamination caused by operational errors.
07
Regular training of experimental personnel
Enhance the aseptic operation awareness and skills of experimental personnel, and conduct relevant training regularly to improve their awareness and ability of contamination prevention.
08
Monitor cell health
The morphology and growth characteristics of the cells are checked regularly, and once abnormalities are found, timely measures should be taken to diagnose and treat possible contamination.
In general, in the face of contamination in cell culture, although there are a range of treatment methods to try to save contaminated cells, the most important thing is to take effective preventive measures to avoid the occurrence of contamination. By strengthening laboratory management, improving operational skills, using high-quality reagent materials and establishing a strict quality control system, the risk of cell contamination can be greatly reduced to ensure the smooth conduct of experiments.
