Laia Ramos1, Javier del Rey1, Gemma Daina1, Olga Martinez-Passarell2, Mariona Rius3, Dolores Tuñón4, Mercedes Campillo5, Jordi Benet1, Joaquima Navarro6. 1. Unitat de Biologia Cel·lular i Genètica Mèdica. Facultat de Medicina, Departament de Biologia Cel·lular, Fisiologia i Immunologia, Universitat Autònoma de Barcelona, Bellaterra, Spain. 2. Fundació Puigvert, Hospital de Sant Pau i de la Santa Creu, Barcelona, Spain. 3. Clínica Sagrada Família, Barcelona, Spain. 4. Institut Universitari Dexeus, Barcelona, Spain. 5. Àrea de Medicina Preventiva i de Salut Pública. Facultat de Medicina. Universitat Autònoma de Barcelona, Bellaterra, Spain. 6. Unitat de Biologia Cel·lular i Genètica Mèdica. Facultat de Medicina, Departament de Biologia Cel·lular, Fisiologia i Immunologia, Universitat Autònoma de Barcelona, Bellaterra, Spain. Electronic address: joaquima.navarro@uab.cat.
Abstract
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.
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.
Authors: Laia Ramos; Javier del Rey; Gemma Daina; Manel García-Aragonés; Lluís Armengol; Alba Fernandez-Encinas; Mònica Parriego; Montserrat Boada; Olga Martinez-Passarell; Maria Rosa Martorell; Oriol Casagran; Jordi Benet; Joaquima Navarro Journal: PLoS One Date: 2014-11-21 Impact factor: 3.240