Match criteria for human cell line authentication: where do we draw the line?

Area of Science:

• Biomedical Research
• Genetics
• Cell Biology

Background:

• Human cell lines are vital research models, but misidentification due to cross-contamination is a significant concern.
• Standardized authentication methods are needed to ensure the reliability of cell line data.
• Short tandem repeat (STR) profiling is recommended for human cell line authentication, but its effectiveness with cultured samples requires assessment.

Purpose of the Study:

• To evaluate the effectiveness of established match criteria for human cell line authentication using STR profiling.
• To assess the reliability of STR profiling in distinguishing between related and unrelated cell lines, including misidentified ones.

Main Methods:

• Analysis of 2,279 STR profiles from four cell banks.
• Grouping 1,157 profiles into sets of related cell lines (duplicates, related, or misidentified).
• Utilizing eight core STR loci plus amelogenin for authentication.

Main Results:

• STR profiling with eight core loci and amelogenin unequivocally authenticated 98% of related cell line sets.
• Two simple match algorithms effectively discriminated between related (≥80% match) and unrelated (<50% match) samples.
• A minor overlap (50-79% match) was observed, primarily with cell lines exhibiting variable STR profiles.

Conclusions:

• The recommended STR profiling match criteria provide a simple and effective method for human cell line authentication.
• Validated criteria enhance the reliability of cell line data in biomedical research.
• Accurate cell line authentication is essential for reproducible scientific outcomes.