Short tandem repeat profiling provides an international reference standard for human cell lines

Abstract

Cross-contamination between cell lines is a longstanding and frequent cause of scientific misrepresentation. Estimates from national testing services indicate that up to 36% of cell lines are of a different origin or species to that claimed. To test a standard method of cell line authentication, 253 human cell lines from banks and research institutes worldwide were analyzed by short tandem repeat profiling. The short tandem repeat profile is a simple numerical code that is reproducible between laboratories, is inexpensive, and can provide an international reference standard for every cell line. If DNA profiling of cell lines is accepted and demanded internationally, scientific misrepresentation because of cross-contamination can be largely eliminated.

Figure 1

Fully analyzed data derived from the DNA of cell line HeLaS3, showing the blue, green, and yellow amplified STR peaks and allele classifications. These data are derived from a single lane containing all the PCR products and the red size standards. The PCR products and size standards are detected by laser excitation of the four fluorescent dye labels by using an Applied Biosystems Prism 377 instrument. Profile quality is assessed by two independent analysts, and the peaks are then assigned allele values corresponding to the number of repeat units by using genescan and genotyper software. Allele designation is based on fixed size windows (±0.5 bp) derived from multiple analyses of allelic ladder samples containing all the commonly occurring alleles (data not shown). The loci are as follows: blue (Left to Right): HUMAMGX/Y, HUMTH01, D21S11, D18S51; green: D8S1179; yellow: HUMVWFA31/A, HUMFIBRA.

Figure 2

Histogram showing relative frequency of the similarity measure at 10% intervals for the 6,903 unrelated pairs of cell lines (black bars) and 264 pairs of cell lines of the same origin (open bars).