Cell contamination is a common problem that plagues every researcher. When using cell culture flasks to culture cells, a small oversight can lead to contamination, negating all previous efforts. Among the various sources of contamination, mycoplasma contamination is one of the most common. The following are general signs of contamination:
Cell shape may not change significantly, mycoplasma can coexist with cells, cell fluids will not die after contamination, and the culture medium generally does not become turbid.
Use of contaminated cells will seriously affect the results of experiments because mycoplasma can inhibit cell growth, cause chromosome abnormalities, alter cell membrane antigens, reduce cell survival rates after recovery, among other effects.
Contamination affects cell metabolism and functionality in the cell culture flask: arginine in the culture medium is consumed in large amounts by mycoplasma, causing a barrier to protein, DNA, rRNA, and mRNA synthesis in cells. Mycoplasma activity causes changes in the composition of the culture medium (e.g., acid production from fermentation-type mycoplasma degrades carbohydrates), which affects cell metabolism.
During culture, more cells break down, the pH of the culture medium changes significantly, and fresh culture medium needs to be replaced frequently.
Cells contaminated with mycoplasma may form a symbiotic system, leading to continued contamination expansion.
There are many reasons for mycoplasma contamination in cell culture flasks, such as cross contamination between cells, contamination from the mouths or skin of operators, contamination of working environments or laboratory equipment, and contamination of culture media. To prevent mycoplasma contamination, strict aseptic operations should be implemented at every step.
Poor cell growth in cell culture flasks is a common problem encountered during experiments, but there is no need to worry too much. If you grasp the factors that affect cell growth, you can plan ahead and ensure that your cells grow happily. As a professional medical oem supplier, Gongdong will introduce you analysis of reasons for poor cell growth in teh cell culture flask.
State of cells
The basic factor affecting future growth is whether the cells are in good condition. If the cells have been passaged too many times, they may be aged. If the inoculation amount is too low, the cell growth rate will be slow; if the cell passage time is too late, the cells will be poisoned, affecting the growth of future passaged cells; if the pancreatic enzyme digestion time is too long or too short, the cells may die if the time is too long, or they may not completely separate and instead become clumped, causing cell death. Slow freezing and rapid thawing of cells for long-term storage and subsequent use may also affect cell growth.
Culture medium or serum
Each type of cell has a suitable serum and culture medium. Selecting or preparing an appropriate culture medium that has not been validated before replacement can also affect cell growth.
Cells require a suitable environment for growth. CO2 supply must be normal, and temperature control by incubators or shakers must be correct.
Contamination by mycoplasma or molds can also affect cell growth.
If the cells in the cell culture flask are not growing well, the reasons can be analyzed from the above aspects to identify the problem and avoid such situations in future experiments.