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Genome-wide uniparental disomy screen in human discarded morphologically abnormal embryos.

Xu J, Zhang M, Niu W, Yao G, Sun B, Bao X, Wang L, Du L, Sun Y - Sci Rep (2015)

Bottom Line: The percentage of UPDs among the morphologically abnormal sourced blastocysts was 3.73%, which is significant higher than the percentage observed in normal blastocysts.The frequency of UPD in 3PN-sourced blastocysts was 7.69%, which is significantly higher than that in normal blastocysts.Our results indicated that UPD may be a common phenomenon in discarded morphologically abnormal embryos and may be relevant to human embryonic self-correction.

View Article: PubMed Central - PubMed

Affiliation: Center for Reproductive Medicine, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, People's Republic of China.

ABSTRACT
Uniparental disomy (UPD) has been shown to be rare in human normal blastocysts, but its frequency in discarded morphologically abnormal embryos and its relevance to embryonic self-correction of aneuploid remains unknown. The aim of this study was to detect UPD in discarded morphologically abnormal embryos. Both discarded morphologically abnormal embryos, including zero-pronuclear zygotes (0PN), one-pronuclear zygotes (1PN), three-pronuclear zygotes (3PN) and 2PN embryos scored as low development potential were cultured into blastocysts then underwent trophectoderm biopsy. Genome-wide UPD screening of the trophectoderm of 241 discarded morphologically abnormal embryo sourced blastocysts showed that UPD occurred in nine embryos. Five embryos exhibited UPDs with euploid chromosomes, and four displayed UPDs with chromosomal aneuploid. The percentage of UPDs among the morphologically abnormal sourced blastocysts was 3.73%, which is significant higher than the percentage observed in normal blastocysts. The frequency of UPD in 3PN-sourced blastocysts was 7.69%, which is significantly higher than that in normal blastocysts. This study provides the first systematic genome-wide profile of UPD in discarded morphologically abnormal embryos. Our results indicated that UPD may be a common phenomenon in discarded morphologically abnormal embryos and may be relevant to human embryonic self-correction.

No MeSH data available.


Comparison of SNP results obtained following the amplification of whole blood DNA and limited cell samples.Multi-cell and limited-cell results indicate the same UPD pattern on these chromosomes. a. UPDs detection on chromosome 1, 7, 12, 13, 19 and 21 using MDA-amplification production of limited cell. b. UPDs detection on chromosome 1, 7, 12, 13, 19 and 21 using global genomic DNA. c. UPDs detection of single cell on chromosome 11, 15 and 20. d. UPDs detection of multi cells on chromosome 11, 15 and 20.
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f1: Comparison of SNP results obtained following the amplification of whole blood DNA and limited cell samples.Multi-cell and limited-cell results indicate the same UPD pattern on these chromosomes. a. UPDs detection on chromosome 1, 7, 12, 13, 19 and 21 using MDA-amplification production of limited cell. b. UPDs detection on chromosome 1, 7, 12, 13, 19 and 21 using global genomic DNA. c. UPDs detection of single cell on chromosome 11, 15 and 20. d. UPDs detection of multi cells on chromosome 11, 15 and 20.

Mentions: To validate the method (using MDA-amplified DNA and SNP array to detect UPD) used in our study, we selected 4 control blood samples and each with 5 replicates for MDA-amplification process. Two normal karyotypes, Arr (1–22) × 2,(XY) × 1, and Arr (1–22) × 2,(X) × 2, were used as negative controls, and two patients with UPD genotypes, [Arr (1–22) × 2, (XY) × 1,UPD(11)(q13.1-q14.1), UPD(15)(q15.1–q21.1),UPD(20)(q13.13-13.33)] and [Arr (1–22) × 2,(X) × 2 UPD(1)(p13.2–p25.1)(q25.3–q32.1),UPD(7)(q35-qter),UPD(12)(q21.2–q23.3),UPD(13)(q12.2–q14.1),UPD(19)(q13.3),UPD(21)(q21.1–q21.3)], were used as positive controls. The G banding karyotypes of these four patients were 46, XX and 46, XY. Genomic DNA was extracted from whole blood samples from each of the four patients for SNP microarray. Simultaneously, to emulate the quantity of cells obtained through trophectoderm biopsy, approximately 3 to 5 peripheral blood lymphocytes were prepared for MDA-amplification. After cell lysis, we used the same genome amplification method as trophectoderm biopsy to perform whole-genome amplification. We performed five repetitions of the amplification procedure and SNP array scan. GenomeStudio software was used to generate SNP genotype calls as previously described for embryos. We acquired the same results using whole blood DNA and limited cell amplification (Figs 1 and 2), which showed that the UPD detection method applied in this study is precise.


Genome-wide uniparental disomy screen in human discarded morphologically abnormal embryos.

Xu J, Zhang M, Niu W, Yao G, Sun B, Bao X, Wang L, Du L, Sun Y - Sci Rep (2015)

Comparison of SNP results obtained following the amplification of whole blood DNA and limited cell samples.Multi-cell and limited-cell results indicate the same UPD pattern on these chromosomes. a. UPDs detection on chromosome 1, 7, 12, 13, 19 and 21 using MDA-amplification production of limited cell. b. UPDs detection on chromosome 1, 7, 12, 13, 19 and 21 using global genomic DNA. c. UPDs detection of single cell on chromosome 11, 15 and 20. d. UPDs detection of multi cells on chromosome 11, 15 and 20.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4508668&req=5

f1: Comparison of SNP results obtained following the amplification of whole blood DNA and limited cell samples.Multi-cell and limited-cell results indicate the same UPD pattern on these chromosomes. a. UPDs detection on chromosome 1, 7, 12, 13, 19 and 21 using MDA-amplification production of limited cell. b. UPDs detection on chromosome 1, 7, 12, 13, 19 and 21 using global genomic DNA. c. UPDs detection of single cell on chromosome 11, 15 and 20. d. UPDs detection of multi cells on chromosome 11, 15 and 20.
Mentions: To validate the method (using MDA-amplified DNA and SNP array to detect UPD) used in our study, we selected 4 control blood samples and each with 5 replicates for MDA-amplification process. Two normal karyotypes, Arr (1–22) × 2,(XY) × 1, and Arr (1–22) × 2,(X) × 2, were used as negative controls, and two patients with UPD genotypes, [Arr (1–22) × 2, (XY) × 1,UPD(11)(q13.1-q14.1), UPD(15)(q15.1–q21.1),UPD(20)(q13.13-13.33)] and [Arr (1–22) × 2,(X) × 2 UPD(1)(p13.2–p25.1)(q25.3–q32.1),UPD(7)(q35-qter),UPD(12)(q21.2–q23.3),UPD(13)(q12.2–q14.1),UPD(19)(q13.3),UPD(21)(q21.1–q21.3)], were used as positive controls. The G banding karyotypes of these four patients were 46, XX and 46, XY. Genomic DNA was extracted from whole blood samples from each of the four patients for SNP microarray. Simultaneously, to emulate the quantity of cells obtained through trophectoderm biopsy, approximately 3 to 5 peripheral blood lymphocytes were prepared for MDA-amplification. After cell lysis, we used the same genome amplification method as trophectoderm biopsy to perform whole-genome amplification. We performed five repetitions of the amplification procedure and SNP array scan. GenomeStudio software was used to generate SNP genotype calls as previously described for embryos. We acquired the same results using whole blood DNA and limited cell amplification (Figs 1 and 2), which showed that the UPD detection method applied in this study is precise.

Bottom Line: The percentage of UPDs among the morphologically abnormal sourced blastocysts was 3.73%, which is significant higher than the percentage observed in normal blastocysts.The frequency of UPD in 3PN-sourced blastocysts was 7.69%, which is significantly higher than that in normal blastocysts.Our results indicated that UPD may be a common phenomenon in discarded morphologically abnormal embryos and may be relevant to human embryonic self-correction.

View Article: PubMed Central - PubMed

Affiliation: Center for Reproductive Medicine, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, People's Republic of China.

ABSTRACT
Uniparental disomy (UPD) has been shown to be rare in human normal blastocysts, but its frequency in discarded morphologically abnormal embryos and its relevance to embryonic self-correction of aneuploid remains unknown. The aim of this study was to detect UPD in discarded morphologically abnormal embryos. Both discarded morphologically abnormal embryos, including zero-pronuclear zygotes (0PN), one-pronuclear zygotes (1PN), three-pronuclear zygotes (3PN) and 2PN embryos scored as low development potential were cultured into blastocysts then underwent trophectoderm biopsy. Genome-wide UPD screening of the trophectoderm of 241 discarded morphologically abnormal embryo sourced blastocysts showed that UPD occurred in nine embryos. Five embryos exhibited UPDs with euploid chromosomes, and four displayed UPDs with chromosomal aneuploid. The percentage of UPDs among the morphologically abnormal sourced blastocysts was 3.73%, which is significant higher than the percentage observed in normal blastocysts. The frequency of UPD in 3PN-sourced blastocysts was 7.69%, which is significantly higher than that in normal blastocysts. This study provides the first systematic genome-wide profile of UPD in discarded morphologically abnormal embryos. Our results indicated that UPD may be a common phenomenon in discarded morphologically abnormal embryos and may be relevant to human embryonic self-correction.

No MeSH data available.