|Alternative Title||Analysis the aneuploidy of frozen-thawed IVF embryos from older women
|Place of Conferral||兰州
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。
|Other Abstract||Objective 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).
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.
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