Does the S phase have an impact on the accuracy of comparative genomic hybridization profiles in single fibroblasts and human blastomeres
Depending on the whole genome amplification used, the cell-cycle stage influences the accuracy of the comparative genome hybridization from single cells, because transitory gains and losses related to DNA replication are detected, such as segmental imbalances.
Laia Ramos, M.D., Javier Del Rey, Ph.D., Gemma Daina, Ph.D., Olga Martinez-Passarell, Ph.D., Mariona Rius, Ph.D., Dolores Tuñón, M.D., Jordi Benet, Professor, Joaquima Navarro, Professor
Volume 101, Issue 2, Pages 488-495.e3, February 2014
To investigate if there is an association between single-cell replicative stage and the segmental chromosome imbalances detected by comparative genomic hybridization (CGH).
First, 135 fibroblasts from cell-line GM03184 (Coriell) at three cell stages (G0/G1, S, and G2/M) were amplified by degenerate oligonucleotide–primed polymerase chain reaction (DOP-PCR) or Sureplex and blindly analyzed by CGH. Second, 85 human blastomeres at the interphase and the metaphase stages, from 30 donated human cryopreserved embryos, were amplified by Sureplex and analyzed by CGH.
Academic center for reproductive medicine.
Main Outcome Measure(s):
Incidence of aneuploidy and segmental imbalances detected at the different cell stages.
In DOP-PCR amplifications of fibroblasts, an increased incidence of segmental abnormalities was detected in the S phase. In Sureplex amplifications of fibroblasts and blastomeres, no differences were detected between the different cell stages. A significantly increased incidence of structural abnormalities was seen in the aneuploid blastomeres.
The segmental imbalances detected after Sureplex amplification in 73.3% of the cryopreserved embryos analyzed are mainly nontransitory. They correspond to segmental imbalances present in the cells due to chromosome instability, rather than to replicative DNA segments.
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