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Mix-ups and mycoplasma: the enemies within
Abstract:
Human leukemia-lymphoma (LL) cell lines represent important tools for experimental research. Among the various problems associated with cell lines, the two most common concern contaminations: (1) cross-contamination with unrelated cells and (2) contamination with microorganisms, in particular mycoplasma. The bad news is that about one-third of the cell lines are either cross-contaminated or mycoplasma-infected or both. The good news is that there are means to recognize and overcome these problems. In cases where, during attempts to establish new LL cell lines, primary LL cultures are cross-contaminated with continuous cell lines, intended new cell lines simply cannot be established ("early" cross-contamination). In cases of "late" cross-contamination of existing LL cell lines where the intrusive cells have a growth advantage, the original ("uncontaminated") cell lines may still be available elsewhere. DNA fingerprinting and cytogenetic analysis appear to be the most suitable approaches to detect cross-contaminations and to authenticate LL cell lines. A different but related aspect of "false" LL cell lines is the frequent misclassification of cell lines whereby the actual cell type of the cell line does not correspond to the purported model character of the cell line. Mycoplasma infection can have a multitude of effects on the eukaryotic cells which, due to the variety of infecting mycoplasma species and many other contributing parameters, cannot be predicted, rendering resulting data questionable at best. Practical procedures for the detection and elimination of mycoplasma contamination have been developed. Diagnostic and preventive strategies in order to hem the alarming increase in "false" and mycoplasma-positive LL cell lines are recommended.
Insights
Human leukemia-lymphoma (LL) cell lines are often contaminated, with about one-third affected by cross-contamination or mycoplasma. DNA fingerprinting and cytogenetic analysis can detect and authenticate LL cell lines, preventing research errors.
Area of Science:
- Cell Biology
- Hematology
- Genetics
Background:
- Human leukemia-lymphoma (LL) cell lines are crucial for research but frequently suffer from cross-contamination with unrelated cells or microbial infections, particularly mycoplasma.
- Approximately one-third of LL cell lines are compromised, potentially leading to inaccurate experimental results.
- Misclassification of cell line identity further complicates their use in research.
Discussion:
- Cross-contamination can occur early during establishment or late in existing cultures, with intrusive cells often having a growth advantage.
- Mycoplasma infections introduce unpredictable effects on eukaryotic cells, compromising data integrity.
- DNA fingerprinting and cytogenetic analysis are effective methods for detecting cross-contaminations and authenticating LL cell lines.
Key Insights:
- Established methods exist for detecting and eliminating mycoplasma contamination.
- Early and late cross-contamination present distinct challenges in LL cell line establishment and maintenance.
- Accurate authentication of LL cell lines is vital to ensure reliable experimental outcomes.
Outlook:
- Implementing diagnostic and preventive strategies is crucial to curb the rise of compromised LL cell lines.
- Further research into robust authentication techniques will enhance the reliability of cell line-based studies.
- Standardized protocols for mycoplasma detection and elimination are recommended for all cell line research.