The use of short tandem repeat profiling to characterize human bladder cancer cell lines.
OBJECTIVE
Cross-contamination of cell lines is a serious but often unrecognized problem. We describe the authentication of a panel of transitional cell carcinoma cell lines using the short tandem repeat profiling technique to detect cross-contamination.


METHODS
Genomic DNA was isolated from UM-UC-1, UM-UC-2, UM-UC-3 (ATCC), UM-UC-6, UM-UC-9, UM-UC-10, UM-UC-11, UM-UC-13, UM-UC-14, UM-UC-16, T24 and KU7 cell lines. Short tandem repeat loci (D3S1358, D16S539, vWA, FGA, TH01, TPOX, CSF1PO, D5S818, D13S317 and D7S820) and a segment of the X-Y homologous gene amelogenin were co-amplified by polymerase chain reaction. Profiling was done using POP-4TM performance optimized polymer 4 (Applied Biosystems) with an ABI Prism 310 genetic analyzer. DNA sequencing of TP53 and immunohistochemistry for p53 were performed in UM-UC-3 and UM-UC-3-GFP.


RESULTS
All cell lines had a unique short tandem repeat profile except UM-UC-2 and T24, which were virtually identical. T24 short tandem repeat profiles matched those of early passage number UM-UC-2. Stable transfection of the green fluorescence protein marker gene did not alter UM-UC-6, UM-UC-14 or KU7 profiles. However, the short tandem repeat profile for UM-UC-3-GFP was different from that of UM-UC-3. DNA sequencing showed a difference in TP53 between UM-UC-3 and UM-UC-3-GFP, confirming that UM-UC-3-GFP is not derived from UM-UC-3.


CONCLUSIONS
Short tandem repeat profiling provides a unique genetic signature of human cell lines that does not significantly change with passage or green fluorescence protein transduction. Using short tandem repeat profiling we noted that the cell line UM-UC-2 is T24. DNA fingerprinting using short tandem repeat profiling is an easy and reliable tool that can be used to verify cell lines.
Chiong
Edmund
E
Department of Urology, University of Texas M. D. Anderson Cancer Center, Houston, Texas 77030, USA.
Dadbin
Ali
A
Harris
Loleta D
LD
Sabichi
Anita L
AL
Grossman
H Barton
HB
D002295:Carcinoma, Transitional Cell / Q000235:genetics* / Q000473:pathology*; D018929:Cell Culture Techniques / Q000379:methods* / Q000592:standards; D045744:Cell Line, Tumor; D002469:Cell Separation / Q000379:methods; D016172:DNA Fingerprinting; D006801:Humans; D018895:Microsatellite Repeats; D001749:Urinary Bladder Neoplasms / Q000235:genetics* / Q000473:pathology*