TRAC 36: Cell Line Identification & Authentication
Laboratories using cultured mammalian cells run the risk of cross-contamination and misidentification of their cell lines. It has been estimated that as many as 20 percent of the cell lines being used for research are not what they are purported to be. Furthermore, the problem continues to grow. Reliable procedures for cell line identification and authentication exist and should be among the important quality control measures practiced by those responsible for the integrity of the research.
After a review of well documented cases and preventative measures, this 3-day lecture and "hands-on" laboratory course will focus on the three most useful methods for authentication and detection of intra- and inter-species cross-contamination. 1. Karyology: basic karyotyping, G and C chromosome banding; marker chromosomes; fluorescent in situ hybridization (FISH); spectral karyotyping. 2. Isoenzymes: genetic and evolutionary basis of isoenzymes; protein/isoenzyme extraction; separation and identification of isoenzyme patterns; application to inter-species identification. 3. DNA Finger-printing/Short Tandem Repeats (STR): genetic and evolutionary basis of DNA fingerprinting; allele selection, detection, and characterization; number of alleles characterized and certitude; DNA profiles/base sequences; multiplex PCR and primer selection; separation by slab gel electrophoresis and capillary electrophoresis; use of automatic sequencers and fluorescent label detection.
Also, cell type identification by intermediate filament immunocytochemistry and proper design of a cell bank will be discussed.
