IDENTIFICATION OF CELL LINES WITH VARIABLE NUMBERS OF TANDEM REPEAT (VNTR) AMPLIFIED BY POLYMERASE CHAIN REACTION

Abstract

We applied VNTR locus typing to the identification of cell lines. Using human leukemia/lymphoma cell lines, each locus (ApoB, D1S80 and DXS52) was amplified by PCR. Detection of theY-chromosome was also done. Sixteen cell lines derived from various individuals, (some cell lines from the same patient, others not) were selected for identification of their ApoB, D1S80and DXS52 VNTR loci and Y-chromosome. BALM-3, -4 and -5, three cell lines derived from one female patient, were identical for all VNTR loci tested. No specific bands for the Ychromosome were detected. BALM-13 and -14, derived from one male patient, were identical for all VNTR loci tested, and their Y-chromosome specific bands were clearly detected. BALM9 and its subclones (BALM-9K, -L, -KL and N), BALM-10, -11 and -12, all derived from one female patient, were as follows: the ApoB locus of BALM-9, B ALM-9KLa nd BALM-12w as detected as two bands of the same size, distinct from the other bands from clones from that patient. The DXS52 locus of BALM-11 was deleted, and BALM-12 showed two bands which were absent in the other clones from the same patient. However, the D1S80 locus was detected consistently in all clones. All results obtained by VNTR locus typing were confirmed by DNA profiling. In further studies, eight human leukemia/lymphoma cell lines, CCRF-HSB-2, MOLT-4, BALL-1, Namalwa, HAL-01, HL-60, KU-812 and EoL-1, were selected and their ApoB, D1S80 and DXS52 loci were investigated at two independent laboratories. The results obtained were compared by computer analysis. They showed identical patterns and bands sizes except for one cell line (Namalwa). This indicated that one cell line from one of the laboratories was mislabeled. We demonstrate here that VNTR typing amplified by PCR is a convenient (simple, speedy and accurate) and useful technique for cell line identification and for checking cross-culture contamination of cell lines.