期刊文献+
共找到2篇文章
< 1 >
每页显示 20 50 100
Immunomagnetic removal of cryo-damaged human spermatozoa 预览
1
作者 UwePaasch SonjaGrunewald KatjaWuendrich TorstenJope Hans-JurgenGlander 《亚洲男性学杂志:英文版》 SCIE CAS CSCD 2005年第1期 61-69,共9页
Aim: To estimate the dissipation of mitochondrial transmembrane potential (mTMP, △ψm) and activation of sperm caspases (aCP) as signs of apoptosis in human spermatozoa during cryopreservation and to evaluate the eff... Aim: To estimate the dissipation of mitochondrial transmembrane potential (mTMP, △ψm) and activation of sperm caspases (aCP) as signs of apoptosis in human spermatozoa during cryopreservation and to evaluate the efficiency of immunomagnetic cell separation (MACS) of these spermatozoa via annexin V-binding. Methods: The mTMP and aCP in fresh and cryopreserved spermatozoa were detected by fluorescence microscopy and by Western blots. The sperm suspensions were divided into two sperm fractions (with intact and deteriorated membranes) by magnetic cell separation (MiniMACS, Miltenyi Biotec, Bergisch Gladbach, Germany) in dependence on their binding to superparamagnetic annexin V-microbeads (AN-MB). Results: The cryopreservation decreased the portion of spermatozoa with intact mTMP from 80.1% ± 7.2 % to 53.5 % ± 13.1% and increased the spermatozoa with activated pancaspases (aCP) from 21.8 % ± 2.6 % to 47.7 % ± 5.8 % (n = 10; mean ± SEM; P < 0.01). The activation of caspases 1, 3, 8, and 9 in the cryopreserved spermatozoa was confirmed by Western blots (n = 22). MACS reduced significantly the percentage of cryopreserved spermatozoa with dissipated mTMP to 8.1 ± 3.9 (P < 0.01) and also those with aCP to 9.3 % ± 2.2 %. Western blot analyses confirmed the increase of the activated caspase3, 9, and 8 in the AN-MB-positive fraction (P < 0.05) compared with the AN-MB-negative fraction. The MACS separation effect was confirmed by anti-annexin V-antibodies. There was no significant influence of the separation column and the magnetic field on the sperm functions. Conclusion: The cryopreservation impaired the mTMP and enhanced the activation status of caspases in human spermatozoa. The immunomagnetic sperm separation via binding of AN-MB could deplete low quality spermatozoa from cryopreserved semen samples. 展开更多
关键词 免疫功能 冷冻损伤 精子 低温贮藏
在线阅读 下载PDF
Stability of fluorochrome based assays to measure subcellular sperm functions 预览
2
作者 Sonja Grunewald Manja Rasch Martin Reinhardt Thomas Baumann Uwe Paasch Hans-Juergen Glander 《亚洲男性学杂志:英文版》 SCIE CAS CSCD 2008年第3期 455-459,共5页
瞄准:为了评估荧光的长期的稳定性,为临床的申请新基于荧光的精液分析试金发信号。方法:从 87 unselected 的精液样品不肥沃的病人被用来执行下列试金:活跃 caspase-3 的(i) 察觉(n= 17 ) ;(i i ) mitochondrial 膜潜力(MMP ) 的正... 瞄准:为了评估荧光的长期的稳定性,为临床的申请新基于荧光的精液分析试金发信号。方法:从 87 unselected 的精液样品不肥沃的病人被用来执行下列试金:活跃 caspase-3 的(i) 察觉(n= 17 ) ;(i i ) mitochondrial 膜潜力(MMP ) 的正直(n= 17 ) ;(iii ) phosphatidylserine (EPS ) 的外表表现(n= 16 ) ;并且(iv ) 经由 CD46 的未经触动的 acrosomes 的察觉(n= 37 ) 。在试金以后, 4% 帕拉甲醛被加到所有 aliquots。每件样品的荧光紧张被流动 cytometry 在天评估 0, 3, 7, 10 和 14。结果:差别直到±,从在白天 0 点测量的价值的 5% 积极精子作为可接受的偏差被估计。Caspase-3 FLICA&#8482;显示出的吝啬的差别 【 5% 在白天 3, 7 和 10 点。在白天 14 点,吝啬的差别是 7.6% 。相反,破坏 MMP 和 EPS 察觉显示出差别 】 5% 在白天 3 点。标记的 CD46-FITC 显示了绝对差别 【 在天的 5%CD46 积极的精子 3, 7, 10 和 14。结论:尽管荧光信号的立即的分析被推荐,在精子染色以后评估 caspase-3 激活直到 10 天和 CD46 直到 14 天是可能的。MMP 和 EPS 察觉的 FACS 评估应该在一样的天被进行。 展开更多
关键词 男性 精子 细胞凋亡 荧光分析
在线阅读 下载PDF
上一页 1 下一页 到第
使用帮助 返回顶部 意见反馈