Cell Line Contamination
Cultured stem cells are vulnerable to contamination by adventitious organisms, such as viruses and bacteria (such as Mycoplasma). Once cell lines are infected, they may undergo spontaneous differentiation or altered function due to adaptation, which may have a profound impact on experimental results. Stem cell laboratories are encouraged to monitor their stem cell cultures. TaqMan® probe-based real-time PCR assays provide a simple and highly sensitive method for detecting specific microorganisms in cell lysates. Compared to traditional methods, such as enzymatic activity or turbidity, TaqMan® Assays offer several advantages, including:
- Sequence-specific PCR primers and probes ensures greater discrimination in detecting target pathogens in complex biological samples.
- Simplified workflow with one standard protocol for multiple organisms.
- Highly sensitive to detect down to 1 colony-forming unit.
- Rapid turnaround time with results in less than 4 hours.
The AuthentiFiler™ PCR Amplification Kit is an effective tool that uses short tandem repeats (STR) genetic loci to confirm the genetic identity of a human cell. As demonstrated by Cabera et. al as well as by other researchers in numerous other publications, STR loci can be an effective way to identify cross contamination events involving two or more human cell lines.
From the leader in real-time PCR, the Applied Biosystems Myco Scan Kit uses real-time PCR to accurately detect Mycoplasma, the most common pathogen infecting high-value cell lines. Often referred to as the “stealth invader” or “silent killer”, Mycoplasma can destroy valuable cell lines and invalidate research results produced using infected cells. Mycoplasma screening is a critical component of the quality control procedures used to protect high-value cell lines such as stem cell lines.
Life Technologies is the leading provider of MycoSEQ Mycoplasma Detection Kits to pharmaceutical companies using cell lines for antibody production and is designed to meet the recommendations set by the European Pharmacopeia for Mycoplasma detection. Now, Myco Scan is available for researchers that want the same high quality results as MycoSEQ, but with a price tag and configuration tailored to research labs.
- Confidence—maintain mycoplasma-free cell lines with the sensitivity and specificity offered only by real-time PCR
- Speed—sample-to-answer in under five hours
- Coverage—detect more than 90 Mycoplasma species with a single assay
Step-by-Step Guide to Cell Line Contamination
Extract nucleic acid from your cells using the protocol of your choice.
Design custom assays for microbial detection or use the MycoSEQ Myco Scan kit for mycoplasma detection.
Applied Biosystems offers more than 1,000,000 TaqMan® Gene Expression Assays, the most comprehensive set of pre-designed Real-Time PCR assays available. All TaqMan Gene Expression Assays have been designed using our validated bioinformatics pipeline, and run with the same PCR protocol, eliminating the need for primer design or PCR optimization.
- Poster: Cardoso, H.A. and Hart, R.P., Establishing standard assays for quality control and identity traceability of human embryonic stem cells (2007)
- White Paper: Eradication of Cross-Contaminated Cell Lines
For research use only. Not intended for any animal or human therapeutic or diagnostic use.