Animal Origin-Free Mycoplasma Removal Reagent

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• Cell Culture ContaminationCell culture contamination falls into six categories: bacteria, yeast, filamentous fungi and moulds, mycoplasma, viruses, and cross-contamination by another cell line. Bacteria, yeast and fungi announce themselves within one to five days through turbidity, a pH shift or visible particles under the microscope, and are handled by discarding the culture and finding the technique failure that let them in. Mycoplasma, viral contamination and cell line misidentification produce no visible change at all and are found only by testing — PCR or DNA stain for mycoplasma, STR profiling for identity. The correct response to any confirmed biological contamination is to discard the affected culture, decontaminate the incubator, and restart from a clean frozen stock.
• Mycoplasma Testing in Cell CultureMycoplasma testing detects Mollicutes species that contaminate laboratory cell cultures without producing turbidity, pH change or any other visible sign, which is why a culture can carry 10⁷–10⁸ organisms per mL and still look healthy. The practical methods are PCR or qPCR (roughly 1–10 CFU/mL, results the same day), enzymatic luminescence assays (about 20 minutes, moderate sensitivity), fluorescent DNA staining read against an indicator cell line (about 10²–10⁶ CFU/mL depending on whether an enrichment step is used), and direct broth-and-agar culture, which remains the reference method but takes up to 28 days. Test cultures on arrival, after quarantine and at least monthly thereafter; a confirmed positive is normally discarded and replaced from a clean frozen stock rather than treated.
• Antibiotics in Cell Culture: Working Concentrations and When Not to Use ThemAntibiotics in cell culture are antibacterial and antifungal agents added to growth medium to suppress microbial contamination; the standard combinations are penicillin–streptomycin at 100 U/mL and 100 µg/mL, gentamicin at 5–50 µg/mL, amphotericin B as an antimycotic at 0.25–2.5 µg/mL, and antibiotic–antimycotic 100X, which supplies all three at 100 U/mL, 100 µg/mL and 0.25 µg/mL when diluted. The professional consensus is that these should not be used routinely or prophylactically: they mask low-level contamination, select for resistant organisms, do nothing at all against mycoplasma, and can alter cell physiology and experimental response. Their defensible uses are primary tissue isolation, work with material from non-sterile sites, and short-term rescue of an irreplaceable culture — not the standard maintenance of established cell lines.