Does the S phase have an impact on the accuracy of comparative genomic hybridization profiles in single fibroblasts and human blastomeres?
Does the S phase have an impact on the accuracy of comparative genomic hybridization profiles in single fibroblasts and human blastomeres?
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DOI：
10.1016/j.fertnstert.2013.10.031
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发表时间：
2014-02-01
影响因子：
6.7
通讯作者：
Navarro, Joaquima
中科院分区：
文献类型：
--
作者：
Ramos, Laia;del Rey, Javier;Navarro, Joaquima
目的：为了研究比较基因组杂交（CGH）检测到的单细胞复制期和节段性染色体不平衡之间是否存在关联，设计：首先，采用简并引物聚合酶链反应（DOP-PCR）或Sureplex对135个细胞系GM 03184（Coriell）的3个细胞期（G 0/G1，S和G2/M）的成纤维细胞进行扩增，并通过CGH进行盲法分析。第二，从30个捐献的人冷冻保存的胚胎中，用Sureplex扩增85个处于间期和中期的人卵裂球，并用CGH分析。地点：生殖医学学术中心。患者：无。干预：无。主要结果测量：在不同细胞阶段检测到的非整倍体和节段不平衡的发生率。结果：在成纤维细胞的DOP-PCR扩增中，在S期检测到节段性异常的发生率增加。在成纤维细胞和卵裂球的Sureplex扩增中，在不同的细胞阶段之间没有检测到差异。结构异常的发生率显着增加被视为在非整倍体blastomeres. Conclusions（S）：后Sureplex扩增73.3%的冷冻保存胚胎分析检测到的节段不平衡主要是暂时的。它们对应于由于染色体不稳定性而存在于细胞中的片段不平衡，而不是复制DNA片段。(C)2014年，美国生殖医学会（American Society for Reproductive Medicine）
Objective: To investigate if there is an association between single-cell replicative stage and the segmental chromosome imbalances detected by comparative genomic hybridization (CGH).Design: 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.Setting: Academic center for reproductive medicine.Patient(s): None.Intervention(s): None.Main Outcome Measure(s): Incidence of aneuploidy and segmental imbalances detected at the different cell stages.Result(s): 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.Conclusion(s): 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. (C) 2014 by American Society for Reproductive Medicine.