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The influence of FSH on the aneuploidy rate of human in vitro matured oocytes
reproductively capable or outside of the designated age range were not eligible for this study. RESULTS: Of the 29 couples contacted 7 individuals participated in this study to date. The questionnaire had a total of 525 possible responses. 24 positve responses were noted (4.6%) and 95.4% were negative. New information regarding genetic risk, miscarriages, learning disabilities, and medication use were identified. CONCLUSIONS: More complete histories were taken with the 3-generation pedigree than the standard family history form. In this study population there were limitations due to small numbers since there was a high non-response rate or refusal. Continuing this study and opening the population to all infertility patients attending this office may affect this outcome. The study is ongoing.
O-46 Monday, October 19, 2009 5:30 PM EPIGENETIC ANALYSIS OF GROWTH AND DEVELOPMENT RELATED IMPRINTED GENES IN HUMAN PLACENTAS DERIVED FROM ASSISTED REPRODUCTIVE TECHNOLOGY. X. Chen, C. Feng, Y. Zhang, D. Zhang, T. Wang, H. Huang. Reproductive Endocrinology, Women’s Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China. OBJECTIVE: To investigate the reasons why the birth weights of assisted reproductive technology (ART) conceived infants are lower than spontaneous conceived infants, even in singletons. DESIGN: We used epigenetic approaches to investigate the differences of growth and development related imprinted genes PHLDA2 and CDKN1C in term and singleton placentas derived from ART and spontaneous pregnancy. MATERIALS AND METHODS: We collected 40 IVF/ICSI conceived, term and singleton placentas and 40 comparative spontaneous conceived placentas. We investigated the methylation patterns for the imprinting control region of PHLDA2 and CDKN1C named KvDMR on human chromosome 11p15 by bisulfite sequencing. We analyzed the monoallelic gene expression of PHLDA2 and CDKN1C by searching SNP and length polymorphism separately. Of particular interest, we also investigated mRNA expression levels of the two genes by real-time PCR. RESULTS: We confirmed that all spontaneous conceived and most ART placentas were differential methylation in KvDMR. However, the metylation patterns of several CpG sites within this region in ART conceived placentas were significantly different from those of spontaneous conceived placentas (p<0.05). Particularly, in one case of ICSI conceived placenta, it was extremely hypomethylation in KvDMR. Both PHLDA2 and CDKN1C were monoallelic expression in placentas and there were no alterations in ART group. In ART conceived placentas CDKN1C mRNA expression levels were significantly upregulated (p<0.05) while there were no significant differences in PHLDA2 expression between two groups. CONCLUSIONS: The present results reveal that abnormal methylation patterns and expressions of growth and development related imprinted genes are involved in ART conceived singleton placentas. It may be one way to explain the lower birth weights of ART conceived singletons. Supported by: No. 30671161 National Natural Science Foundation of China.
and 22 by multi-color fluorescence in situ hybridization. The effect of FSH on spindle morphology and chromosome alignment in metaphase II oocytes were then explored using immunocytochemistry. RESULTS: Totally, 136 oocytes were matured in vitro and 131 were successfully biopsied. Clear hybridization signals were observed on 120 fixed polar bodies and oocytes respectively. Overall, 87.5% of the diagnoses based on the polar bodies were in accordance with the results on the corresponding oocytes. The aneuploidy rates were 26.7, 23.3, 36.7 and 63.3% in four groups respectively. Significantly higher aneuploidy rate was detected in oocytes matured in the group with the highest FSH concentration (26 IU/ml) (P¼0.008). No differences in gross spindle morphology and chromosome configuration were obeserved between high (26IU/ml) and conventional (0.075IU/ml) FSH concentration groups. CONCLUSIONS: High FSH concentration in IVM medium significantly increased the first meiotic division error, resulting in more aneuploidy oocytes during in vitro maturation. Polar body biopsy combined with FISH is an effective approach for oocyte aneuploidy screening.
O-48 Monday, October 19, 2009 6:00 PM STUDY ON THE INCIDENCE OF CHROMOSOMAL ABNORMALITIES IN 10325 INFERTILITY PATIENTS WHO RESORT TO IVF/ ICSI. P. Liang, Y. Zeng, B. Yin, Q. Lin, J. Cai, W. Zhang. Shenzhen Zhongshan Urology Hospital ART Center, Shenzhen Zhongshan Institute for Reproductive Medicine and Genetics, Shenzhen, Guangdong, China. OBJECTIVE: To determine the prevalence of chromosomal abnormalities in 10325 IVF/ICSI patients. DESIGN: Retrospective study. MATERIALS AND METHODS: The peripheral blood were collected from the infertility couples who resort to IVF/ICSI in our IVF center respectively. Chromosome specimens were acquired by routine culture of peripheral blood lymphocytes and G band technique was used. RESULTS: 658 cases with abnormal karyotype were detected in 10325 IVF/ICSI patients, the incidence of abnormal karyotype was 6.37%. The incidence rates of different types of abnormal karyotype and the cases of abnormal chromosome showed clinical characters related to reproduction refer to TABLE 1
The type of abnormal karyotype Chromosome heteromorphism
The cases detected The ratio The incidence in 10325 in 658 rate in 10325 IVF/ICSI abnormal IVF/ICSI patients karyotype patients 498
75.68%
4.82%
Sex chromosomal abnormalities
55
8.36%
0.533%
Balanced translocation
49
7.45%
0.474%
Mosaic
25
3.80%
0.242%
Robertsonian translocation
20
3.04%
0.194%
Pericentric inversion
6
0.91%
0.058%
Others
8
1.22%
0.077%
O-47 Monday, October 19, 2009 5:45 PM THE INFLUENCE OF FSH ON THE ANEUPLOIDY RATE OF HUMAN IN VITRO MATURED OOCYTES. P. Y. Ting, X. Y. Wen. Reproductive Medical Center, First Affiliated Hospital of Sun Yat-sen University, Guang Zhou, Guang Dong, China. OBJECTIVE: 1,To study the effect of Follicle-Stimulating Hormone (FSH) on the aneuploidy rate of human oocytes during in vitro maturation. 2,To study the influence of FSH on the spindle morphology of human oocytes matured in vitro. 3,To establish preimplantation genetic aneuploidy screening (PGS) test by chromosome analysis of the first polar body (1PBs). DESIGN: In vitro model study. MATERIALS AND METHODS: Immature oocytes, which were discarded in intracytoplasmic sperm injection (ICSI) cycles, were collected from Oct. 2008 to May 2009 and randomly allocated into four groups according to FSH concentrations (0, 0.075IU/ml, 0.3 IU/ml, 26IU/ml) in IVM medium. Polarization microscopy was used for spindle visualization.Fixed polar bodies and corresponding oocytes were tested on chromosomes 13, 16, 18, 21
S14
Abstracts
The cases of abnormal chromosome showed clinical characters related to reproduction Oligozoospermia (16 cases), Spontaneous abortion(8 cases). Azoospermia(24 cases), Oligozoospermia and asthenozoospermia (15 cases), Turner’s Syndrome(5 cases). Azoospermia (2 cases), Oligozoospermia (9 cases), Asthenozoospermia (1 cases), Spontaneous abortion (10 cases), Recurrent miscarriage (7 cases), Terata (2 cases), amenorrhea (1 cases). Oligozoospermia (1 cases), Spontaneous abortion (3 cases). Oligozoospermia (2 cases), Spontaneous abortion (5 cases), Recurrent miscarriage (1 cases). Spontaneous abortion (3 cases), Recurrent miscarriage (1 cases). Oligozoospermia (2 cases), Spontaneous abortion (1 cases).
Vol. 92., No. 3, Supplement, September 2009
O-46 Monday, October 19, 2009 5:30 PM EPIGENETIC ANALYSIS OF GROWTH AND DEVELOPMENT RELATED IMPRINTED GENES IN HUMAN PLACENTAS DERIVED FROM ASSISTED REPRODUCTIVE TECHNOLOGY. X. Chen, C. Feng, Y. Zhang, D. Zhang, T. Wang, H. Huang. Reproductive Endocrinology, Women’s Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China. OBJECTIVE: To investigate the reasons why the birth weights of assisted reproductive technology (ART) conceived infants are lower than spontaneous conceived infants, even in singletons. DESIGN: We used epigenetic approaches to investigate the differences of growth and development related imprinted genes PHLDA2 and CDKN1C in term and singleton placentas derived from ART and spontaneous pregnancy. MATERIALS AND METHODS: We collected 40 IVF/ICSI conceived, term and singleton placentas and 40 comparative spontaneous conceived placentas. We investigated the methylation patterns for the imprinting control region of PHLDA2 and CDKN1C named KvDMR on human chromosome 11p15 by bisulfite sequencing. We analyzed the monoallelic gene expression of PHLDA2 and CDKN1C by searching SNP and length polymorphism separately. Of particular interest, we also investigated mRNA expression levels of the two genes by real-time PCR. RESULTS: We confirmed that all spontaneous conceived and most ART placentas were differential methylation in KvDMR. However, the metylation patterns of several CpG sites within this region in ART conceived placentas were significantly different from those of spontaneous conceived placentas (p<0.05). Particularly, in one case of ICSI conceived placenta, it was extremely hypomethylation in KvDMR. Both PHLDA2 and CDKN1C were monoallelic expression in placentas and there were no alterations in ART group. In ART conceived placentas CDKN1C mRNA expression levels were significantly upregulated (p<0.05) while there were no significant differences in PHLDA2 expression between two groups. CONCLUSIONS: The present results reveal that abnormal methylation patterns and expressions of growth and development related imprinted genes are involved in ART conceived singleton placentas. It may be one way to explain the lower birth weights of ART conceived singletons. Supported by: No. 30671161 National Natural Science Foundation of China.
and 22 by multi-color fluorescence in situ hybridization. The effect of FSH on spindle morphology and chromosome alignment in metaphase II oocytes were then explored using immunocytochemistry. RESULTS: Totally, 136 oocytes were matured in vitro and 131 were successfully biopsied. Clear hybridization signals were observed on 120 fixed polar bodies and oocytes respectively. Overall, 87.5% of the diagnoses based on the polar bodies were in accordance with the results on the corresponding oocytes. The aneuploidy rates were 26.7, 23.3, 36.7 and 63.3% in four groups respectively. Significantly higher aneuploidy rate was detected in oocytes matured in the group with the highest FSH concentration (26 IU/ml) (P¼0.008). No differences in gross spindle morphology and chromosome configuration were obeserved between high (26IU/ml) and conventional (0.075IU/ml) FSH concentration groups. CONCLUSIONS: High FSH concentration in IVM medium significantly increased the first meiotic division error, resulting in more aneuploidy oocytes during in vitro maturation. Polar body biopsy combined with FISH is an effective approach for oocyte aneuploidy screening.
O-48 Monday, October 19, 2009 6:00 PM STUDY ON THE INCIDENCE OF CHROMOSOMAL ABNORMALITIES IN 10325 INFERTILITY PATIENTS WHO RESORT TO IVF/ ICSI. P. Liang, Y. Zeng, B. Yin, Q. Lin, J. Cai, W. Zhang. Shenzhen Zhongshan Urology Hospital ART Center, Shenzhen Zhongshan Institute for Reproductive Medicine and Genetics, Shenzhen, Guangdong, China. OBJECTIVE: To determine the prevalence of chromosomal abnormalities in 10325 IVF/ICSI patients. DESIGN: Retrospective study. MATERIALS AND METHODS: The peripheral blood were collected from the infertility couples who resort to IVF/ICSI in our IVF center respectively. Chromosome specimens were acquired by routine culture of peripheral blood lymphocytes and G band technique was used. RESULTS: 658 cases with abnormal karyotype were detected in 10325 IVF/ICSI patients, the incidence of abnormal karyotype was 6.37%. The incidence rates of different types of abnormal karyotype and the cases of abnormal chromosome showed clinical characters related to reproduction refer to TABLE 1
The type of abnormal karyotype Chromosome heteromorphism
The cases detected The ratio The incidence in 10325 in 658 rate in 10325 IVF/ICSI abnormal IVF/ICSI patients karyotype patients 498
75.68%
4.82%
Sex chromosomal abnormalities
55
8.36%
0.533%
Balanced translocation
49
7.45%
0.474%
Mosaic
25
3.80%
0.242%
Robertsonian translocation
20
3.04%
0.194%
Pericentric inversion
6
0.91%
0.058%
Others
8
1.22%
0.077%
O-47 Monday, October 19, 2009 5:45 PM THE INFLUENCE OF FSH ON THE ANEUPLOIDY RATE OF HUMAN IN VITRO MATURED OOCYTES. P. Y. Ting, X. Y. Wen. Reproductive Medical Center, First Affiliated Hospital of Sun Yat-sen University, Guang Zhou, Guang Dong, China. OBJECTIVE: 1,To study the effect of Follicle-Stimulating Hormone (FSH) on the aneuploidy rate of human oocytes during in vitro maturation. 2,To study the influence of FSH on the spindle morphology of human oocytes matured in vitro. 3,To establish preimplantation genetic aneuploidy screening (PGS) test by chromosome analysis of the first polar body (1PBs). DESIGN: In vitro model study. MATERIALS AND METHODS: Immature oocytes, which were discarded in intracytoplasmic sperm injection (ICSI) cycles, were collected from Oct. 2008 to May 2009 and randomly allocated into four groups according to FSH concentrations (0, 0.075IU/ml, 0.3 IU/ml, 26IU/ml) in IVM medium. Polarization microscopy was used for spindle visualization.Fixed polar bodies and corresponding oocytes were tested on chromosomes 13, 16, 18, 21
S14
Abstracts
The cases of abnormal chromosome showed clinical characters related to reproduction Oligozoospermia (16 cases), Spontaneous abortion(8 cases). Azoospermia(24 cases), Oligozoospermia and asthenozoospermia (15 cases), Turner’s Syndrome(5 cases). Azoospermia (2 cases), Oligozoospermia (9 cases), Asthenozoospermia (1 cases), Spontaneous abortion (10 cases), Recurrent miscarriage (7 cases), Terata (2 cases), amenorrhea (1 cases). Oligozoospermia (1 cases), Spontaneous abortion (3 cases). Oligozoospermia (2 cases), Spontaneous abortion (5 cases), Recurrent miscarriage (1 cases). Spontaneous abortion (3 cases), Recurrent miscarriage (1 cases). Oligozoospermia (2 cases), Spontaneous abortion (1 cases).
Vol. 92., No. 3, Supplement, September 2009