兰州大学机构库 >学院待认领
高育龄妇女体外受精冻融胚胎非整倍体检测的研究
Alternative TitleAnalysis the aneuploidy of frozen-thawed IVF embryos from older women
宋北剑
Thesis Advisor张学红
2011-05-25
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name硕士
Keyword非整倍体 荧光原位杂交 染色体异常 植入前遗传学诊断
Abstract目的 利用FISH技术,选用13,18,21,X,Y五色荧光探针,研究高龄妇女IVF受精后冻融胚胎的非整倍体分布。 方法 收集IVF-ET治疗后的冷冻胚胎,纳入标准为:①科研胚胎,患者经IVF-ET或FET治疗后均已成功妊娠并分娩正常胎儿,夫妻双方签署知情同意书,同意将剩余冷冻胚胎用于科学研究;②女方因素所致的不孕,且女方年龄≥35岁;③双方染色体均无明显异常。共收集来自于7对夫妇43枚胚胎,所有胚胎解冻后采用OCTAX-LaserShot激光透明带打孔,吸拉法活检每个胚胎中所有卵裂球,使用Tween-20/HCI+甲醇/冰醋酸法固定每个胚胎的所有检出卵裂球,选用MultiVysion PGT探针(包括13、18、21、X、Y五色荧光探针)对所固定的卵裂球行FISH后进行分析。 结果 收集来自7对夫妇43枚胚胎,解冻后成功复苏40枚胚胎,复苏率为93.02%。2枚胚胎因存活细胞数≤1而排除。共41枚胚胎270个存活卵裂球,活检后,获得完整卵裂球264个,活检率为97.78%(264/270)。264个卵裂球中有核卵裂球243个(92.05%),12个卵裂球含双核,2个卵裂球含三核,共有259个核,成功固定251个核,固定率为96.91%(251/259)。杂交后247个核有信号,杂交率为98.41%(247/251)。 41枚胚胎中,9枚为正常胚胎,32枚为异常胚胎,其中异常非嵌合型胚胎2枚,无规律分离型胚胎5枚,嵌合型胚胎25枚。二倍体嵌合型胚胎17枚,异常嵌合型8枚。一致性胚胎11枚,非一致性胚胎30枚。 11枚胚胎含有多核细胞,均为异常胚胎。14个多核卵裂球共30个核,其中22个核为异常,占73.33%,明显高于单核组(P<0.05)。 41个胚胎共247个核有杂交信号,其中103个核为异常,占41.70%。在13、18、21、X、Y五种染色中,18号染色体出现异常的频率最高,其次是13号和21号染色体,性染色体的异常频率最低。 随着胚胎细胞数目的增多,二倍型嵌合型胚胎内的染色体异常细胞的比例逐渐下降(rs= -0.520,P<0.05)。 结论 1. 高育龄妇女IVF受精后冻融胚胎中,染色体异常广泛存在,并且主要以嵌合形式存在。 2. 在13,18,21,X,Y五中染色体中,18号染色体在高育龄妇女IVF受精后冻融胚胎中的异常频率最高,其次为13、21号,性染色体异常频率最低。 3. 随着胚胎细胞数目的增多,二倍型嵌合胚胎中染色体异常细胞的比例逐渐下降。胚胎发育过程中可能存在一定的自我修复机制。 4. 多核细胞核的染色体异常率明显高于单核细胞。
Other AbstractObjective To detect the aneuploidy of the frozen-thawed IVF embryos from older women by FISH with fluorescent probes contained chromosomes 13, 18, 21, X and Y. Methods The frozen embryos after IVF-ET treatment were collected. Inclusion criteria: ①Patients had been successful pregnancy and delivery after IVF-ET or FET treatment, and the couples had signed consent form with the intention to donate their remaining frozen embryos for scientific research; ② Female infertility factors with the women age ≥35 years; ③There was no obvious chromosomal abnormalities with the couples. A total of 43 embryos from 7 couples were selected. All blastomeres of each embryo were acquired through biopsy after drilling a hole on the zona pellucida by OCTAX-LaserShot. The blastomeres were fixed using Tween-20/HCI + methanol/acetic acid. Fluorescence in situ hybridization processed with MultiVysion PGT probes (including chromosomes 13, 18, 21, X, Y five-color fluorescent probes). Then the FISH signals were analysised. Results A total of 43 embryos from 7 couples were selected .40 embryos were successfully revived after thawed. The recovery rate was 93.02%. 2 embryos were excluded because in which there was only 1 alive cell. A totel of 270 blastomeres were alive in 41 embryos. 264 intact blastomeres (97.78%) were acquired after biopsy. 243 blastomeres(92.05%) in 264 blastomeres had nucleus,of which 12 blastomere had two nuclei and 2 blastomeres had 3 nuclei. 251 of 259(96.91%) nuclei were fixed successfully. 247 of 251(98.41%) nuclei had FISH signals. 9 embryos out of 41 were uniformly normal. 32 embryos were abnormal, of which 2 non-mosaic embryos, 5 chaotic embryos, 25 mosaic embryos. There were 17 diploid mosaic embryos and 8 abnormal mosaic embryos embryos. 11 embryos were uniform embryos, 30 embryos were non-uniform embryos. 11 embryos out of 41 had cells with multinuclei. All of those embryos were abnormal. 22 out of 30 nuclei (73.33%) from 14 blastomeres were abnormal in cells with multinuclei. The abnormal rate was significantly higher than single-nuclei group (P <0.05). 103 out of 247 nuclei (41.70%) from 41 embryos nucleis were abnormal. The abnormal frequency of the chromosome 18 was the highest in the five chromosomes, followed by chromosome 13 and 21. The abnormal frequency of the sex chromosome was the lowest. In the diploid mosaic embryos, with the number of embryonic cells increased, the proportion of abnormal cells was decreased (rs = -0.520, P< 0.05). C...
URL查看原文
Language中文
Document Type学位论文
Identifierhttps://ir.lzu.edu.cn/handle/262010/223439
Collection学院待认领
Affiliation临床医学院
Recommended Citation
GB/T 7714
宋北剑. 高育龄妇女体外受精冻融胚胎非整倍体检测的研究[D]. 兰州. 兰州大学,2011.
Files in This Item:
There are no files associated with this item.
Related Services
Recommend this item
Bookmark
Usage statistics
Export to Endnote
Altmetrics Score
Google Scholar
Similar articles in Google Scholar
[宋北剑]'s Articles
Baidu academic
Similar articles in Baidu academic
[宋北剑]'s Articles
Bing Scholar
Similar articles in Bing Scholar
[宋北剑]'s Articles
Terms of Use
No data!
Social Bookmark/Share
No comment.
Items in the repository are protected by copyright, with all rights reserved, unless otherwise indicated.