一种稳定的人卵母细胞染色体荧光原位杂交技术

A refined technique for fluorescence in situ hybridization with human oocyte chromosomes

目的:建立一种稳定、简便的人卵母细胞染色体荧光原位杂交技术,为揭示母方减数分裂错误及其发生原因、探讨非整倍性发生机制、评价外环境因素对人卵母细胞的遗传学效应提供有力工具。方法:经签订知情同意书后,取体外受精术后废弃的人卵母细胞,经低渗、渐进固定处理、制备人卵母细胞染色体核型。应用人16号染色体着丝粒荧光探针,经变性处理后与人卵母细胞核型进行荧光原位杂交,再经洗脱和染色等步骤,在荧光显微镜下分析。结果:经过上百批次重复实验,人卵母细胞染色体形态与分散良好,荧光信号明亮清晰、背景干净。结论:成功建立人卵母细胞染色体荧光原位杂交技术。

OBJECTIVE： In humans,aneuploidies occur in at least 5% of all clinically recognized pregnancies,among which about 90% can be caused by maternal meiotic error. In this study,a stable and simple method for fluorescence in situ hybridization（FISH） with human oocyte chromosomes was developed to explore causes of maternal meiotic error,mechanisms of aneuploidy and genetic effects of environmental factors on human oocytes. METHODS： The abandoned human oocytes were collected from IVF patients who signed consent forms. The oocytes were used to prepare chromosomal karyotype through hypotonic and gradual fixative treatments. Human oocyte metaphase FISH was performed using the CEP 16 Probe. RESULTS：From our observations,the morphology and dispersal of human oocyte chromosomes were excellent,and the fluorescence signals were bright and clear with clean background. CONCLUSION：We established successfully the method for FISH with human oocyte chromosomes.