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The most common source is through contamination of cultures with microorganisms such as bacteria, fungi and yeasts, which can usually be observed under the microscope.
Another important contaminant is mycoplasma, which is resistant to traditional cell culture sterilisation techniques, isn’t visible and requires specific routine testing to detect. Once a mycoplasma contamination has been detected the only way to ensure a mycoplasma free environment is through hydrogen peroxide fumigation, also known as “bombing”, of the lab. All cell lines from Abcam are supplied mycoplasma-free, so this is something you should not need to worry about.
Aseptic technique prevents contamination of cell culture and reagents from microorganisms in the environment. This video will outline the procedures necessary to work in a sterile manner within a cell culture hood.
Before commencing work ensure that you have wash your hands and are wearing the correct personal protective equipment, this includes gloves a lab coat and safety glasses. Thoroughly spray the inside of the cell culture hood with disinfectant and wipe clean with tissue.
Dispose of used tissue in the appropriate waste bin. Next spray the inside of the hood with 70% ethanol and wipe clean with tissue. The hood is now ready to use. Take the cell culture medium and supplements from the fridge and place in a 37oC incubator to warm. To prepare the medium aseptic spray each item with ethanol. Aim to work quickly and efficiently to minimize the risk of contamination and place each bottle in easy reach. Add each supplement to the bottle of medium using a new sterile tip or pipette. Only open bottles immediately before use and if the lid is put down insure the opening is facing upwards. Never sweep your hands and sleeves over the top of an opened bottled as this could cause contamination. To minimize the risk of this happening replace the lid as soon as you are finished with the bottle.
Label the medium bottle with your initials the date and details of the contents.