Syauqy Ahmad, Umar Liya Agustin, Lestari Silvia Werdhy, Mansyur Eliza, Hestiantoro Andon, Paradowszka-Dogan Agnieszka
a Department of Medical Biology, Faculty of Medicine , University of Indonesia , Jakarta , Indonesia.
b Master Program in Biomedical Sciences, Faculty of Medicine , University of Indonesia , Jakarta , Indonesia.
Syst Biol Reprod Med. 2016 Oct;62(5):309-16. doi: 10.1080/19396368.2016.1210695. Epub 2016 Aug 3.
This study aimed to evaluate sperm chromatin maturity and integrity of that injected into good-quality oocytes in an in vitro fertilization-intra cytoplasmic sperm injection (IVF-ICSI) program. A cut-off value of sperm chromatin maturity and integrity was developed as a function of their correlation to the zygote development, i.e., embryo formation and cleavage rate. The study assessed sperm chromatin maturity using aniline blue (AB) staining, whereas toluidine blue (TB) staining was used to assess sperm chromatin integrity. Ejaculates from 59 patients undergoing ICSI and 46 fertile normozoospermic donors for determination of normal values of sperm chromatin status were used in this study. Embryo formation and cleavage rates were observed for the period of 3 days after ICSI. There was a significant difference in the percentage of sperm with mature chromatin between ejaculate from ICSI patients and fertile donor (p=0.020); while there was no significant difference in sperm chromatin integrity of both samples (p=0.120). There was no significant correlation between sperm chromatin maturity and either embryo formation or cleavage rate; as well as sperm chromatin integrity to both parameters of zygote development (p>0.05). Furthermore, we found that the cut-off value of sperm chromatin maturity and integrity of the fertile normozoospermic ejaculates were 87.2% and 80.2%, respectively. Using the cut-offs, we found that low sperm chromatin maturity at the level of <87% correlated significantly with the cleavage rate of the zygote (p=0.022; r=0.371); whereas poor sperm chromatin integrity at the level of <80% correlated with embryo formation (p=0.048; r=0,485). In conclusion, this study showed that poor maturity and integrity of sperm chromatin (AB<87% and TB<80%, respectively), could affect zygote development following ICSI.
AB: aniline blue; CMA3: chromomycin A3; ICSI: intra cytoplasmic sperm injection; IVF: in vitro fertilization; PBS: phosphate buffer saline; SPSS: Statistical Package for Social Science; TB: toluidine blue; WHO: World Health Organization.
本研究旨在评估体外受精 - 卵胞浆内单精子注射(IVF - ICSI)程序中注入优质卵母细胞的精子染色质成熟度和完整性。根据精子染色质成熟度和完整性与合子发育(即胚胎形成和分裂率)的相关性,制定了一个截断值。本研究使用苯胺蓝(AB)染色评估精子染色质成熟度,而使用甲苯胺蓝(TB)染色评估精子染色质完整性。本研究使用了59例接受ICSI治疗的患者以及46例生育力正常的正常精子捐献者的精液来确定精子染色质状态的正常值。在ICSI后3天观察胚胎形成和分裂率。ICSI患者精液与生育力正常捐献者精液中染色质成熟精子的百分比存在显著差异(p = 0.020);而两个样本的精子染色质完整性无显著差异(p = 0.120)。精子染色质成熟度与胚胎形成或分裂率之间无显著相关性;精子染色质完整性与合子发育的两个参数之间也无显著相关性(p>0.05)。此外,我们发现生育力正常的正常精子精液中精子染色质成熟度和完整性的截断值分别为87.2%和80.2%。使用这些截断值,我们发现染色质成熟度低于87%与合子的分裂率显著相关(p = 0.022;r = 0.371);而染色质完整性低于80%与胚胎形成相关(p = 0.048;r = 0.485)。总之,本研究表明精子染色质成熟度和完整性较差(分别为AB<87%和TB<80%),可能会影响ICSI后的合子发育。
AB:苯胺蓝;CMA3:放线菌素A3;ICSI:卵胞浆内单精子注射;IVF:体外受精;PBS:磷酸盐缓冲盐水;SPSS:社会科学统计软件包;TB:甲苯胺蓝;WHO:世界卫生组织。