Figure 7
Genomic integration of cfCh in vitro and in vivo analyzed by FISH and PCR. (a) Representative FISH images showing presence of abundant human whole-genomic signals in metaphase spreads prepared from NIH3T3 cells co-cultivated with dead Jurkat cells at tenth passage (upper panel). No signals were detected when live Jurkat cells were similarly used. The human whole-genomic probe used did not cross react with mouse DNA. Quantitative analysis of number of human signals per metaphase (lower panel). Thirty metaphases were analyzed in each case. The figure represents mean values (±S.E.). (b) Gel image of PCR-amplified products showing human pan Alu elements in clones E7 and B10. Upper panel, lane 1: 1 kb ladder; lane 2: NIH3T3 cells (negative control); lane 3: human DOK cells (positive control); lane 4 and 5: E7 and B10 clones (respectively); lane 6: no template control. Lower panel, input control PCR, mouse β-ACTIN (104 bp) and human HER2 gene (136 bp) were amplified in mouse NIH3T3 clones and human DOK cells, respectively. (c) Detection of human DNA signals by FISH in vital organs of mice following i.v. injection of dead and live Jurkat cells and their prevention by concurrent treatment with CNPs, DNase I and R-Cu. One hundred thousand Jurkat cells were injected i.v. and animals killed after 7 days. Two animals were used in each case and 500 nuclei were examined for each organ. Percent cells showing human signals were recorded and mean values were compared by Χ2-test. **P<0.01, ****P<0.0001. The human whole-genomic probe used did not cross react with mouse DNA.
