The need for the identification of cell lines is critical before starting an experiment or research. There are incidents where research and experimentations had been withdrawn after decades of work after realizing that a given cell line was misidentified at the source. On other occasions, the cells either got genetically drifted or cross-contaminated with time. Finally, the scientific faculty realized the signification of Cell Line Authentication.
A minimum of 80% match between the STR profiles of the cell line under study and the reference cell line is standard. However, further investigations are required to authenticate the originality of the cell line of interest.
Common Problems
Inarguably, the misidentification of a given cell line is the most common and critical problem. When the cells are collected at the source, their accurate identification is vital for the success of the experiment or the research. Misidentified at this point, the research/experiment may go on for years and even decades without providing a conclusive or accurate result.
One of the most commonly observed problems with the cell lines can be a complete loss of identity, meaning the cells of a given cell line may become completely different cells over time. These changes can be triggered by factors like the culture medium, wherein the cells are stored, or other environmental conditions.
Another common problem is genetic drift. The passage of time or the passage of a cell line from the test animals (like that of the mice) may cause the loss of heterozygosity, leading to genetic drifting, thus changing the genetic identity.
Cross-contamination of the cell lines, with the environmental pollutants, microorganisms, etc., is also a problem geneticists and researchers frequently face.
The Procedure of Cell Line Authentication Test
A Cell Line Authentication test can accurately detect misidentified, genetically drifted, and cross-contaminated cell lines. The procedure used for Cell Line Authentication is STR profiling of the cell line under study and matching it with the STR profile of the reference cell line.
The main steps involved in a Cell Line Authentication Test are:
• Collecting the cell of the cell-line under investigation.
• Extracting the DNA from the cell.
• Analyzing the DNA’s STR regions having polymorphism (Short Tandem Repeats).
• PCR (Polymerase Chain Reaction) amplification of the polymorphic sites.
• Capillary Electrophoresis of the PCR products.
• Profile creation of the repeating regions.
Where to Get an Accurate & Reliable Cell Line Authentication Test in India?
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