We performed DNA fingerprinting to verify that TiPS shared a genetic background with the starting donor T-cell population and to rule out cell line cross-contamination (Figure S1).
In view of this unexpected result, DNA fingerprinting of HeLa and HT1080 vector and hTR cells was used to confirm that there had been no cell line cross-contamination with ALT cells (data not shown).
These include the cracks on the skin, entering through the eyes, which might be particularly vulnerable because of the limited activity of the immune system (McKenna and Chen, 2010), and inhalation of aerosols, which are commonly formed while handling cells and have been documented as a route of cell line cross-contamination (Torsvik et al, 2010).
Huh7 cell line authentication and intraspecies cell line cross-contamination were analyzed using Promega's StemElite ID System (Madison, WI, USA) for the following short tandem repeats (STRs): D21S11, TH01, TPOX, vWA, CSF1PO, D16S539, D7S820, D13S317 and D5S818; Amelogenin was also analyzed for gender identification together with a specific marker for mouse DNA.
Cell-line cross-contamination was excluded using short tandem repeat profiling, as described.
"Health and safety issues, such as bacterial infection, cross contamination excessive line speeds and exploitation, such as low wages, all create a climate that strongly mitigates against best practice," says Singh, who added that the problems were compounded by a largely migrant workforce with poor training and language difficulties.
Furthermore, the origin of several of the cell lines has been questioned; in several instances STR profiling has revealed cross contamination or misidentification of established cell lines, including several GBM lines (Capes-Davis et al., 2010).
Here we describe a comprehensive and cost-efficient SNP-based solution to the problems of mouse cell line misidentification, cross-contamination and copy number aberration.
STR analysis allowed to exclude possible cross contamination of analysed cell lines excluding H-318 (murine cell line).
The finding of the identical mutation raised the suspicion of cross contamination between cell lines.
Separate dosing lines and leak valves were used for water to avoid cross contamination and accidental exposure of TMA to water in the dosing manifold.
