Esmaeilpour Tahereh, Elyasi Leila, Bahmanpour Soghra, Ghannadi Alireza, Monabbati Ahmad, Dehghani Farzaneh, Kazerooni Marjaneh
Department of Anatomical Sciences, Medical School, Shiraz University of Medical Sciences, Shiraz, Iran.
Islamic Azad University, Larestan, Iran. ; Shiraz Infertility Center, Shiraz, Iran.
Iran J Reprod Med. 2012 Sep;10(5):435-40.
It has been claimed that by using different washing methods, the sperms can be separated according to size, motility, density, chromosomal content and surface markings and charge. These methods also reduce sperm chromatin deficiencies and screen the sperms before applying in assisted reproduction techniques.
This study compared simple density gradient methods and a combined method with albumin density gradient and PureSperm separation (alb/PureSperm) for sex preselection by double fluorescence in situ hybridization (FISH) versus chromomycin A3 staining to determine chromatin integrity.
30 normal semen samples were prepared with PureSperm, albumin gradients and alb/PureSperm. All samples were then stained by FISH and chromomycin A3. The results were compared with SPSS 11.5 and the Kruskal-Wallis test.
The proportion of X-bearing spermatozoa by PureSperm separation (47.58±5.67) and Y-bearing spermatozoa by albumin gradient (46.13±3.83) methods were slightly higher than in putative normal sperm samples (1:1), but there were no significant differences in the X- or Y- bearing spermatozoa counts among the three methods. Albumin gradient separation tended to underestimate abnormal spermatozoa compared to PureSperm and combined alb/PureSperm.
Routine separation methods slightly enriched X- or Y- bearing spermatozoa, but the differences were not significant for clinical purposes. The combined alb/PureSperm method had no advantages for assessing sex ratio or chromatin integrity compared to simpler gradient methods.
据称，通过使用不同的洗涤方法，可以根据精子的大小、活力、密度、染色体含量以及表面标记和电荷对精子进行分离。这些方法还能减少精子染色质缺陷，并在应用于辅助生殖技术之前对精子进行筛选。
本研究比较了简单密度梯度法与白蛋白密度梯度和PureSperm分离法联合使用（alb/PureSperm），通过双重荧光原位杂交（FISH）与嗜铬霉素A3染色进行性别预选，以确定染色质完整性。
用PureSperm、白蛋白梯度和alb/PureSperm制备30份正常精液样本。然后所有样本均用FISH和嗜铬霉素A3染色。结果采用SPSS 11.5和Kruskal-Wallis检验进行比较。
通过PureSperm分离法得到的含X精子比例（47.58±5.67）和通过白蛋白梯度法得到的含Y精子比例（46.13±3.83）略高于假定的正常精子样本（1:1），但三种方法在含X或含Y精子计数上无显著差异。与PureSperm和联合alb/PureSperm法相比，白蛋白梯度分离法往往会低估异常精子。
常规分离方法能略微富集含X或含Y精子，但就临床目的而言差异不显著。与更简单的梯度法相比，联合alb/PureSperm法在评估性别比例或染色质完整性方面没有优势。
