(1) How to avoid cell contamination
The types of cell contamination can be classified into bacteria, fungi, molds, viruses, and the like. The main causes of contamination are improper aseptic technique, poor operating room environment, contaminated serum and contaminated cells. Strict aseptic technique, clean environment, and good quality cell source and media formulation are the best ways to reduce contamination.
(2) What is the effect of mycoplasma-contaminated cells on cell culture?
The abnormality of mycoplasma contamination cannot be observed with the naked eye. Except for highly experienced experts, most cell lines contaminated with mycoplasma cannot be distinguished by their appearance.
Mycoplasma contamination can affect almost all cell growth parameters, metabolism and study any data. Therefore, before performing the experiment, it must be confirmed that the cells are mycoplasma-free, and the data of the experimental results are meaningful.
(3) The medium is stored in a refrigerator at 4 °C, the color will be dark red, and the pH will become more alkaline.
The medium is stored in a refrigerator at 4 °C, and the CO 2 in the medium will gradually overflow, causing the medium to become more alkaline, and the color of the acid-base indicator (usually phenol red) in the medium will also vary with the alkali. Increased sex and more dark red. As a result of the alkaline medium, the cell growth will be stagnant or dead. If the medium is alkaline, the CO 2 can be sterile filtered to adjust the pH. Or adjust the pH with acid and alkali.
(d) Is L-glutamine important in cell culture? Is it unstable in solution?
L-glutamine is important in cell culture. After the amino group is removed, L-glutamine can be used as a source of energy for cultured cells, involved in protein synthesis, and nucleic acid metabolism. L-Glutamine degrades over time in solution, and degradation of L-glutamine leads to the formation of ammonia, which is toxic to some cells.
(5) What medium can be omitted to add phenol red? What is the role of sodium pyruvate in the medium?
Phenol red is used as an indicator of pH in the medium: red in neutral, yellow in acidity, and purple in alkaline. Due to phenol red interference detection, some researchers did not use phenol red-containing medium for flow cytometry.
Sodium pyruvate can be used as an alternative carbon source in cell culture. Although cells are more likely to use glucose as a carbon source, cells can also metabolize sodium pyruvate without glucose.
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