Cell banks are crucial to biologics production, providing consistent, well-characterized cell lines for manufacturing. However, mycoplasma contamination can occur during cell culture due to raw materials or handling, making routine testing essential for product safety and regulatory compliance.
Cefety Bioscience conducts mycoplasma testing according to USP, EP, and Chinese Pharmacopoeia standards. We utilize both the culture method and the indicator cell culture method, which together form the gold standard for reliable detection.
The culture method involves inoculating samples into validated liquid and solid media, with regular passaging and observation. While highly sensitive, this method may miss strains that do not form visible colonies.
To address this, the indicator cell method co-cultures the sample with indicator cells, followed by fluorescent DNA staining and microscopic observation to detect even hard-to-culture mycoplasma.
Culture Method
Highly sensitive and widely accepted, the culture method involves:
Inoculating test samples into both liquid and solid nutrient media.
Including negative and positive controls (e.g., Mycoplasma pneumoniae ATCC 15531, Mycoplasma orale ATCC 23714).
Performing serial passaging on days 2, 7, 14, and 21.
Monitoring for colony formation or growth inhibition.
Indicator Cell Culture Method
This method complements the culture assay by detecting difficult-to-culture mycoplasma:
1.Co-culturing test samples with Vero indicator cells in six-well plates.
2.Including relevant controls such as Mycoplasma hyorhinis and Mycoplasma orale.
3.Passaging and staining with fluorescent DNA dyes.
4.Microscopic observation for mycoplasma fluorescence on cell surfaces.
Note: Prior to testing, it's essential to confirm that test samples do not contain substances that inhibit mycoplasma growth, to ensure accurate results.
By combining traditional culture and indicator cell assays-the gold standard in mycoplasma detection-Cefety Bioscience offers reliable, highly sensitive testing that meets global regulatory requirements and supports the safe production of biologics. This method is implemented in strict accordance with USP 2024, EP 11.0, and the Chinese Pharmacopoeia 2020 Edition, ensuring full compliance with international standards and consistent detection of mycoplasma contamination.
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