I'm trying to start a fish embryonic cell line. The cells are growing up and dividing but the bacterium multiply and kill cells. We do everything aseptic but the grow and colonise our cultures.
We usual use Streptomycin, Penicillin and Kanamycin but one or two days after that, we realise the became die, they start multiply.
According to my experience working with human umbilical vein endothelial cells (HUVECs) and endothelial progenitor cells (EPCs), the contamination with bacteria will release toxins in media which will change the cellular metabolism, thus leading to their great decrease and apoptosis. Indeed, penicillin and streptomycin are strong antibiotics. Those antibiotics are one of the components of the growth media I use in my experiments. Another antibiotic you may use for further culture is 0.2% Normocin. However, you will waste so much time trying to remove contamination from your culture and you will never know exactly what happened. In addition, there is a great possibility that a small number of bacteria will survive and contaminating your cells. Finally, you run the risk of contaminating the cells from other workers by keeping these cells in the incubator.
I would like to tell you that the most scientifically based option is to start the culture again!!! You must clean and sterilize very well your culture room and the incubator. You must also prepare new clean buffers and solutions and check media are free of bacteria. All the workers in the lab should use right cell culture techniques and keep culture lab clean. When everything is ok, take a vial of cells from the nitrogen and put them in a plate to grow. Finally, if you want to check whether your medium is free of bacteria or not, you put only growth medium in a sterilized plate without adding cells. Then, you put it in the incubator at 37o C and examine after 1 day the colour of your medium. If it will change and it is different, it means that it is contaminated.
I hope that I helped you!!
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