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Environmental Exposure of Sperm Sex-Chromosomes: A Gender Selection Technique

机译:精子性染色体的环境暴露:性别选择技术

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摘要

Preconceptual sex selection is still a highly debatable process whereby X- and Y-chromosome-bearing spermatozoa are isolated prior to fertilization of the oocyte. Although various separation techniques are available, none can guarantee 100% accuracy. The aim of this study was to separate X- and Y-chromosome-bearing spermatozoa using methods based on the viability difference between the X- and Y-chromosome-bearing spermatozoa. A total of 18 experimental semen samples were used, written consent was obtained from all donors and results were analysed in a blinded fashion. Spermatozoa were exposed to different pH values (5.5, 6.5, 7.5, 8.5, and 9.5), increased temperatures (37°C, 41°C, and 45°C) and ROS level (50 μM, 750 μM, and 1,000 μM). The live and dead cell separation was done through a modified swim-up technique. Changes in the sex-chromosome ratio of samples were established by double-label fluorescent in situ hybridization (FISH) before and after processing. The results indicated successful enrichment of Xchromosome-bearing spermatozoa upon incubation in acidic media, increased temperatures, and elevated H2O2. This study demonstrated the potential role for exploring the physiological differences between X-and Y-chromosome-bearing spermatozoa in the development of preconceptual gender selection.
机译:避孕前的性别选择仍然是一个值得商process的过程,通过该过程,在卵母细胞受精之前分离出带有X和Y染色体的精子。尽管可以使用各种分离技术,但没有一种可以保证100%的准确性。本研究的目的是使用基于X和Y染色体的精子活力差异的方法分离X和Y染色体的精子。总共使用了18个实验精液样品,所有供体均获得了书面同意,并以盲法分析了结果。精子暴露于不同的pH值(5.5、6.5、7.5、8.5和9.5),升高的温度(37°C,41°C和45°C)和ROS水平(50μM,750μM和1,000μM) 。活细胞和死细胞的分离通过改良的泳动技术完成。在处理之前和之后,通过双标记荧光原位杂交(FISH)确定样品的性染色体比率的变化。结果表明,在酸性介质中孵育,温度升高和H2O2升高后,富含X染色体的精子得以成功富集。这项研究表明,在概念前性别选择的发展中,探索X和Y染色体精子之间生理差异的潜在作用。

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