How much do you know about Mycoplasma contamination in cells?
Nowadays, everything from biopetroleum to medicines, medical care, etc. is closely related to life science. Cell culture is the most basic experimental technology in the entire life science research. The development and application of stem cell biology and vaccine research and development are largely based on the development of cell culture technology.
Cell culture is to disperse cells from tissues or primary explant blocks, so as to be suspended, and then culture them on a solid matrix to grow adherently, or to be suspended in culture medium.
cell lines are an important material for life science research, and zero-contamination cell lines are the prerequisite for obtaining reliable experimental data. The sources of microorganisms contamination in generally include bacteria, fungi, and mycoplasma . Among them, the Mycoplasma contamination rate is as high as 30% to 60%. Part of the reason why mycoplasma contamination is so common is that it can spread effectively even in conventional subculture .
1. What is Mycoplasma
Mycoplasma, also known as Mold, has a diameter of 0.1-0.3μm. It is the smallest and simplest prokaryotic found so far. Mycoplasma can be easily passed through the filter membrane and mixed into the culture system, showing a highly versatile and has a variety of forms such as spherical, rod-shaped, filamentous, and branch-shaped. Usually attached to the cell membrane surface, Mycoplasma does not have rigid cell wall , so ordinary antibiotics are ineffective against it.
Common types in the laboratory include: Mycoplasma oral, Mycoplasma porcine nasal, Mycoplasma fermentation, Mycoplasma humanoid, Mycoplasma salivary, Mycoplasma pulmonary and Mycoplasma piloma, etc.
2. Consequences of Mycoplasma contamination
cell lines once the cell line is infected with Mycoplasma, the culture medium usually does not have color changes. Mycoplasma attaches to the surface of the cell line and proliferates by seizing nutrients from the cell culture medium. In addition to consuming various nutrients in the culture medium during its reproduction, it also changes the pH value in the culture medium, and even produces metabolites that are toxic to the cell line, resulting in changes in the cell state. Although the cell line will not die immediately after contamination of
Mycoplasma, it will affect various parameters in the cell, such as changing cell membrane antigenicity, cell metabolism, interfering with nucleic acid synthesis, changing the efficiency of DNA transfection, leading to chromosomal aberration, etc., which has far-reaching effects in many aspects and seriously affecting the accuracy of various parameters of cultured cells.
III. Source of Mycoplasma contamination
Potential ways of Mycoplasma contamination include:
Contaminated culture medium, reagent or instrument, including sterile reagents and materials, laminar flow ultra-clean workbench, constant temperature and humidity incubator, water bath pot and refrigerator, etc.
· Laboratory personnel (about 80.6% of the technicians are Mycoplasma carriers)
· Infected cultures in other laboratories are contaminated by multiple Mycoplasmas at the same time under laboratory culture strips. This is a problem that most laboratories have a headache. Staying aseptic is a huge challenge for both newbies and veterans. Cells cultured in vitro lack an immune system, so the way to control pollution is to prevent pollution! The traditional disinfection process of cell laboratory is: 2. Turn on the ultra-clean workbench and cell room, and irradiate for 1~2h. It is worth noting that after ultraviolet light is turned on, people cannot enter the laboratory, and ultraviolet rays are harmful to both eyes and exposed skin. To enter, the operator must turn off the UV light first and turn it on again after the person leaves. cell Mycoplasma contamination solution
1. Wipe the countertop with disinfectant alcohol;
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