Cell Line STR Presentation

Human Cell Line Quality Control Through STR Analysis At ATCC

By Greg Sykes, Robyn Hedges, Ming Hui, and Yvonne Reid, ATCC

ABSTRACT: The ATCC Cell Biology Department conducts quality control tests on human cell lines, including isoenzymology, morphological examinations, and microbial contamination checks. Part of accessioning a new human cell line involves a Short Tandem Repeat (STR) analysis either to authenticate the uniqueness of the sample or to confirm a relationship to cell lines sharing a common origin. The profile is also checked for cross-contamination with other cell lines. The analysis is conducted using the PowerPlex 1.2 kit by Promega, which was originally optimized for the identification of normal human specimens, with applications primarily in forensics and paternity tests. Upon receiving a new human cell line from a depositor, a portion is spotted and archived on FTA paper. The cell line is expanded into master cell bank (seed) and working cell bank (distribution) stocks. The baseline profile generated from the master stock—if not from earlier material—is first compared against all other baselines stored in an in-house database. The working stock, which is prepared from the master cell bank stock, is made available to the research community. Every human cell line produced by the ATCC must pass STR analysis (match the baseline profile) prior to distribution. Reproducible data from human cell lines are generated by the PowerPlex 1.2 kit, but unlike normal human material, cell lines typically yield unbalanced peaks and may express a loss or gain of alleles. Chromosomal shuffling and displacements, uneven karyokinesis, and genetic mutations associated with cancer and experimentally altered cells are major contributors to these profile anomalies. Variations, such as a loss of heterozygocity, are sometimes found in cancer tissue profiles when compared to normal tissue data from the same patient. Understanding the facts surrounding cell culture profiles and the use of a commercially available identification kit enables researchers worldwide to verify existing laboratory specimens and to communicate cell line authenticity.

This seminar was presented at the American Chemical Society conference in May 2002. The audience consisted of fellow scientists already familiar with STR technology and how it applies to forensics, paternity testing, and the like. If STRs are new to you, NIST has an excellent introductory page called Brief Introduction to STRs. Once you have some STR background, click on this page’s PowerPoint presentation, Human Cell Line Quality Control Through STR Analysis At ATCC. For further descriptions, open the presentation and in the PowerPoint menu bar, go to “View,” “Notes Page” to see the notes on each slide.

Steaming ampoule (pulled from liquid nitrogen dewar) and building photography by G.R. SYKES.