STR DNA Profiling: The Standard for Cell Line AuthenticationJun 05, 2014 at 12:00 PM ET
Misidentification of human cell lines is a common problem that leads to irreproducible research in the Life Sciences. Over the years, use of contaminated cell lines has increased due to poor techniques, inadequate authentication protocols, and sharing of false cell lines among researchers. These concerns can be readily resolved by the implementation of the Standard STR protocol–authentication of human cell lines by STR profiling. In this webinar, we will discuss the recent advances in STR profiling technologies and will delve into further detail on how the Standard STR protocol is transforming scientific practices.
Douglas R. Storts, PhD
Head of Research - Nucleic Acid Technologies, Promega Corporation
Yvonne Reid, PhD
Yvonne A. Reid, Ph.D., is a Consultant and former employee at ATCC. Dr. Reid has been at the forefront of developing and standardizing methods to characterize cell lines by enzymology, karyotyping, and more recently DNA profiling, and to propagate, cryopreserve, and track validated batches of thousands of animal cell lines and strains. These validation protocols have been adopted by cell banks around the world. In addition, Dr. Reid has been heavily involved in developing guidelines for authenticating cell lines and cell types.
Can the COI primer sets be shared for the different ATCC species that ATCC uses, or is there a reference for these primer sets?
Reference: Species identification in cell culture: a two-pronged molecular approach. Jason K Cooper, Greg Sykes, Steve King, Karin Cottrill, Natalia V Ivanova, Robert Hanner, Pranvera Ikonomi, In Vitro Cellular & Developmental Biology - Animal, 2007; 43(10):344- 51
Can you identify tagged cell lines?
The GenePrint and PowerPlex Systems only amplify a defined set of STR loci and a gender marker (amelogenin). Consequently, they cannot distinguish a tagged cell line from an untagged cell line unless the tag is inserted into the loci being amplified.
If the subclone of one cell line has a characteristic trisomy so that one allele is doubled, then is the ratio between alleles as seen by STR analysis consistent enough to be used to differentiate the two?
It is possible to distinguish cell lines by relative peak height; however, the peak heights may vary from experiment-to-experiment. Consequently, it is preferable to identify unique alleles that can be used to distinguish the subclones.
What is your recommended STR profiling frequency for pure research applications?
Cell lines should be genotyped upon receipt into the laboratory, unless they are sourced from a reputable supplier that has already confirmed the identity of the cell line. Identity should also be confirmed prior to submitting the manuscript for publication. If the laboratory uses multiple cell lines, or the project duration is long, there may be merit to performing genotyping on occasion to avoid wasting valuable research effort using the wrong cell line.