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Characterization of rainbow trout cell lines using microsatellite DNA profiling.
G M Perry1, G J McDonald, M M Ferguson
1Department of Zoology, University of Guelph, Guelph, N1G 2W1, Canada.
Cytotechnology
|November 13, 2008
Summary
Genetic profiling using ten microsatellite loci confirmed the uniqueness of salmonid cell lines. This method effectively identified a misidentified rainbow trout cell line as Chinook salmon, highlighting its utility in preventing cross-contamination.
Area of Science:
- Molecular Biology
- Aquaculture Genetics
- Cell Line Authentication
Background:
- Maintaining authentic cell lines is crucial for reproducible research in aquaculture and fisheries.
- Genetic drift and cross-contamination can compromise the integrity of established cell lines.
- Microsatellite markers offer a powerful tool for distinguishing closely related individuals and cell lines.
Purpose of the Study:
- To develop and apply a multiplexed microsatellite genotyping assay for salmonid cell line authentication.
- To establish genetic profiles for various rainbow trout (Oncorhynchus mykiss) and Chinook salmon (O. tschawytscha) cell lines.
- To validate the utility of microsatellite analysis in identifying cell line misidentification and cross-contamination.
Main Methods:
- Amplification of ten specific microsatellite loci using the polymerase chain reaction (PCR).
- Genotyping of nine rainbow trout cell lines, one Chinook salmon cell line, and a Pacific herring (Clupea harengus pallasi) control.
- Comparative analysis of genetic profiles across all tested cell lines to assess uniqueness.
Main Results:
- All tested cell lines, except for two derived from the same fish (RTP-91E and RTP-91F), showed unique genetic profiles.
- The microsatellite locus Ots1BML proved most effective in differentiating between cell lines.
- An uncharacterized rainbow trout cell line (Clone 1A) was definitively identified as CHSE-214 (Chinook salmon).
Conclusions:
- Multiplexed microsatellite analysis is a reliable and efficient method for creating genetic profiles of salmonid cell lines.
- This approach is highly effective for verifying cell line identity and detecting cross-contamination.
- Accurate cell line authentication is essential for ensuring the validity of research findings in fish cell culture.
