Study of mitochondrial membrane potential, reactive oxygen species, DNA fragmentation and cell viability by flow cytometry in human sperm
Study of mitochondrial membrane potential, reactive oxygen species, DNA fragmentation and cell viability by flow cytometry in human sperm
复制标题
DOI：
10.1093/humrep/17.5.1257
复制
发表时间：
2002-05-01
影响因子：
6.1
通讯作者：
Marchetti, P
中科院分区：
文献类型：
--
作者：
Marchetti, C;Obert, G;Marchetti, P
背景：精子细胞死亡似乎是男性不育的一个原因。这项研究的目的是确定在体外受精精子准备过程中评估精液样本中精子质量的最可靠方法。方法：对精液样本进行常规分析，并与几种检测死亡相关变化的细胞荧光方法进行比较。对不育症患者的纯净精液和PureSperm(R)梯度法制备的精子进行常规显微镜观察和线粒体膜电位(Delapsi(M))、活性氧生成、DNA片段化和细胞存活率分析。结果：在纯净精液中，高精细胞率与标准精液参数(浓度/活力)呈正相关。受精率高的精子细胞和DNA碎片低的精子细胞在体外受精后显示高受精率。在准备好的精子中可以检测到的唯一变化是Delapsi(M)的变化，Delapsi(M)(高)精子与前向活动率和体外受精后的高受精率呈正相关。结论：线粒体膜电位分析是判断精子质量最敏感的检测方法。这些发现有望开发出一种有助于预测试管受精成功的测试方法。
BACKGROUND: Sperm cell death appears to be a cause of male infertility. The objective of this study was to determine the most reliable method for the evaluation of sperm quality in semen samples during sperm preparation for IVF. METHODS: Conventional analysis of semen samples was compared with several cytofluorometric methods detecting death-associated changes. Neat semen from infertile patients and sperm prepared by PureSperm((R)) gradient were studied by conventional microscopy and analysed for mitochondrial membrane potential (Deltapsi(m)), generation of reactive oxygen species, DNA fragmentation and cell viability. RESULTS: In neat semen, a positive correlation was found between the percentage of Deltapsi(m)(high) sperm cells and standard semen parameters (concentration/motility). Sperm cells depicting Deltapsi(m)(high) and cells with low DNA fragmentation displayed high fertilization rate after IVF. The only changes that could be detected in prepared sperm were changes in Deltapsi(m), with Deltapsi(m)(high) sperm positively correlated with forward motility and also with high fertilization rates after IVF. CONCLUSION: Analysis of mitochondrial membrane potential is the most sensitive test by which to determine sperm quality. These findings promise development of a test that may help to predict successful IVF.