Abstract
Objective To evaluate the effectiveness of several methods in eliminating Mycoplasma contamination in cell culture of human hepatoma-derived cell line C3A. Methods PCR was performed to detect the Mycoplasmas contamination in cell cultures. The contaminated samples were treated by ciprofloxacin, heating, Plasmocure or co-culturing with macrophages. Transmission electron microscope (TEM) and Q-PCR were used to comparatively analyze the cell morphology and gene expression before and after Plasmocure treatment. Results Plasmocure succeeded in eliminating Mycoplasma contamination, while ciprofloxacin showed temporary efficacy. Heating and co-culturing with macrophages failed to eliminate Mycoplasma contamination. No Mycoplasma contamination in the Plasmocure-treated group was observed under TEM and the expression of ALB, TF and CYP3A4 genes were higher than the genes expressed in the contaminated group (P<0.01). Conclusion Plasmocure treatment was effective in eliminating Mycoplasma contamination in cell culture. Moreover, the cell morphology and gene expression in Plasmocure-treated group were restored to normal. Key words: Mycoplasma contamination; Elimination method; Cell culture
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