The use of mammalian cell culture in the study of human cellular physiological processes has been demonstrated. Microbial contamination, on the other hand, is still a prevalent and important problem for researchers around the world, and it happens even when strict aseptic procedures are followed. This can have major consequences, such as corrupted results and the loss of crucial biological samples due to erroneous, non-reproducible data. Mycoplasma is the most prevalent causal agent found in mammalian cell lines. The identification and eradication of bacterial contamination observed during mammalian cell line culture are described in this article. Because of the contamination, we were on the verge of losing the cell lines. Using 16S rDNA sequencing, the contaminant was identified as gram-positive bacteria from the species Bacillus that was already resistant to penicillin and streptomycin. The cultures were treated for two weeks with a cocktail of antibiotics, including ciprofloxacin and gentamicin, in varied doses to determine the most efficient curing dose with the least cytotoxicity. The cells were eventually found to be bacterial-free. As a result, even in the absence of additional antimicrobial treatment, cell lines maintained for an extended period of time (under the microscope) remained healthy and normal. Finally, using FACS, we discovered that these cells preserved their typical cell cycle features.
Author (s) Details
Ms. Garima Singh
Molecular Biology Unit, Institute of Medical Sciences, Banaras Hindu University, Varanasi, UP- 221005, India.
Ms. Kumud Tiwari
Molecular Biology Unit, Institute of Medical Sciences, Banaras Hindu University, Varanasi, UP- 221005, India.
Mr. Samarendra K. Singh
Molecular Biology Unit, Institute of Medical Sciences, Banaras Hindu University, Varanasi, UP- 221005, India and School of Biotechnology, Institute of Science, Banaras Hindu University, Varanasi, UP- 221005, India.
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