人类异常受精卵及胚胎的染色体组成
发布时间：2019-03-18 13:47
【摘要】： 目的: 采用单细胞染色体制备技术,分析人类常规体外受精与胚胎移植中(in-vitro fertilization and embryo transfer, IVF-ET )异常受精卵及异常受精卵来源胚胎的染色体组成。 方法: 加精后24～72小时收集异常受精卵75枚、异常受精卵来源的胚胎277枚,其中IVF来源的三原核(tripronuclear, 3PN)受精卵71枚、2细胞胚胎38枚、2细胞胚胎239枚,卵胞浆内单精子显微注射(intracytoplasmic sperm injection,ICSI)来源的单原核(monopronucleus, 1PN)胚胎4枚。分别与秋水仙素(终浓度0.10 ug/ml)共培养14～20小时,Tyrode’s液去除透明带,胚胎需分散成卵裂球,0.9%枸橼酸钠液低渗20～30分钟后梯度固定,胰酶显带、Giemsa染色,美国维士染色体核型自动分析系统(Auto-System K)分析核型并照相。 结果: (1) IVF-3PN受精卵及胚胎的染色体制备:3PN受精卵的有丝分裂指数及染色体制备成功率最高,分别为67.6% (48/71)、52.1% (37/71)。2细胞胚胎的有丝分裂指数及染色体制备成功率分别为38.2%(29/76)、31.2%(24/76)。2细胞胚胎的有丝分裂指数及染色体制备成功率最低,分别为16.2%(149/920)、8.2%(75/920)。 (2) 3PN受精卵及胚胎的染色体组成:3PN受精卵的三倍体率最高,为75.7%(28/37),2细胞胚胎为70.8%(17/24),2细胞胚胎染色体核型紊乱,多为近二倍体(56%)、近单倍体(32%),三倍体率仅为9.3%(7/75)。 (3) ICSI-1PN胚胎的染色体组成:4枚ICSI-1PN胚胎分散后共获29枚细胞,其中6细胞(来自于3枚胚胎)可染色体计数,显示均为单倍体(5细胞为23条染色体,1细胞为25条染色体)。 结论: (1)传统染色体制备方法可对受精卵及胚胎行全套染色体检测,但技术操作难度大、染色体制备成功率低,该方法尚需进一步改进。 (2) IVF-3PN受精卵及-2细胞胚胎多为三倍体,2细胞胚胎多为近二倍体、近单倍体,其染色体核型紊乱。 (3) ICSI-1PN胚胎多为单倍体。 (4) IVF-3PN及ICSI-1PN胚胎均不能用于移植。
[Abstract]:Aim: to analyze the chromosome composition of abnormal fertilized eggs and embryos derived from abnormal fertilized eggs during conventional in vitro fertilization and embryo transfer (in-vitro fertilization and embryo transfer, IVF-ET) by single-cell chromosome preparation technique. Methods: 75 abnormal fertilized eggs and 277 abnormal fertilized eggs were collected 24 hours after fertilization, including 71 IVF-derived tripronuclear (tripronuclear, 3PN) fertilized eggs, 38 2-cell embryos and 239 2-cell embryos. Four monopronucleus, 1PN embryos derived from intracytoplasmic sperm microinjection of (intracytoplasmic sperm injection,ICSI were obtained. Colchicine (final concentration 0.10 ug/ml) was co-cultured with colchicine for 14 hours for 20 hours. The clear zone was removed by Tyrode' 's solution. The embryos were dispersed into blastomeres. After 20 minutes of hypotonic solution of 0.9% sodium citrate for 30 minutes, gradient fixation and trypsin banding were observed. Giemsa staining and automatic karyotype analysis system (Auto-System K) were used for karyotype analysis and photography. Results: (1) chromosome preparation of IVF-3PN fertilized eggs and embryos: mitotic index and successful rate of chromosome preparation of 3PN fertilized eggs were 67.6% (48 / 71), respectively. The mitotic index and the success rate of chromosome preparation were 38.2% (29 / 76) and 38.2% (29 / 76), respectively. The mitotic index and chromosome preparation success rate of the 2-cell embryos were the lowest, 16.2% (149 / 920) and 8.2% (75 / 920), respectively. (2) chromosome composition of 3PN fertilized eggs and embryos: the triploid rate of 3PN fertilized eggs was 75.7% (28 / 37), that of 2-cell embryos was 70.8% (17 / 24), and the karyotype of 2-cell embryos was abnormal. Most of them were near diploid (56%), near haploid (32%) and triploid rate was only 9.3% (7 / 75). (3) chromosome composition of ICSI-1PN embryos: after 4 ICSI-1PN embryos were dispersed, 29 cells were obtained, of which 6 cells (from 3 embryos) could be counted, and all of them were haploid (5 cells were 23 chromosomes). 1 the cells were 25 chromosomes. Conclusion: (1) the traditional chromosome preparation method can be used to detect the whole set of chromosomes in fertilized eggs and embryos, but the technical operation is difficult and the success rate of chromosome preparation is low. This method needs to be further improved. (2) IVF-3PN fertilized eggs and-2-cell embryos were mostly triploid, 2-cell embryos were nearly diploid and nearly haploid, and their karyotypes were disordered. (3) ICSI-1PN embryos were haploid. (4) both IVF-3PN and ICSI-1PN embryos could not be used for transplantation.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R321
本文编号：2442922
[Abstract]:Aim: to analyze the chromosome composition of abnormal fertilized eggs and embryos derived from abnormal fertilized eggs during conventional in vitro fertilization and embryo transfer (in-vitro fertilization and embryo transfer, IVF-ET) by single-cell chromosome preparation technique. Methods: 75 abnormal fertilized eggs and 277 abnormal fertilized eggs were collected 24 hours after fertilization, including 71 IVF-derived tripronuclear (tripronuclear, 3PN) fertilized eggs, 38 2-cell embryos and 239 2-cell embryos. Four monopronucleus, 1PN embryos derived from intracytoplasmic sperm microinjection of (intracytoplasmic sperm injection,ICSI were obtained. Colchicine (final concentration 0.10 ug/ml) was co-cultured with colchicine for 14 hours for 20 hours. The clear zone was removed by Tyrode' 's solution. The embryos were dispersed into blastomeres. After 20 minutes of hypotonic solution of 0.9% sodium citrate for 30 minutes, gradient fixation and trypsin banding were observed. Giemsa staining and automatic karyotype analysis system (Auto-System K) were used for karyotype analysis and photography. Results: (1) chromosome preparation of IVF-3PN fertilized eggs and embryos: mitotic index and successful rate of chromosome preparation of 3PN fertilized eggs were 67.6% (48 / 71), respectively. The mitotic index and the success rate of chromosome preparation were 38.2% (29 / 76) and 38.2% (29 / 76), respectively. The mitotic index and chromosome preparation success rate of the 2-cell embryos were the lowest, 16.2% (149 / 920) and 8.2% (75 / 920), respectively. (2) chromosome composition of 3PN fertilized eggs and embryos: the triploid rate of 3PN fertilized eggs was 75.7% (28 / 37), that of 2-cell embryos was 70.8% (17 / 24), and the karyotype of 2-cell embryos was abnormal. Most of them were near diploid (56%), near haploid (32%) and triploid rate was only 9.3% (7 / 75). (3) chromosome composition of ICSI-1PN embryos: after 4 ICSI-1PN embryos were dispersed, 29 cells were obtained, of which 6 cells (from 3 embryos) could be counted, and all of them were haploid (5 cells were 23 chromosomes). 1 the cells were 25 chromosomes. Conclusion: (1) the traditional chromosome preparation method can be used to detect the whole set of chromosomes in fertilized eggs and embryos, but the technical operation is difficult and the success rate of chromosome preparation is low. This method needs to be further improved. (2) IVF-3PN fertilized eggs and-2-cell embryos were mostly triploid, 2-cell embryos were nearly diploid and nearly haploid, and their karyotypes were disordered. (3) ICSI-1PN embryos were haploid. (4) both IVF-3PN and ICSI-1PN embryos could not be used for transplantation.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R321
【参考文献】
相关期刊论文 前1条
1 叶虹,黄国宁,裴莉,陈春秀,刘红;促性腺激素释放激素激动剂长、短方案在体外受精胚胎移植中的比较性研究[J];中华妇产科杂志;2001年04期
，本文编号：2442922
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