Tetrazolium Salt WST-8 as a Novel and Reliable Chromogenic Indicator for the Assessment of Boar Semen Quality.
Tetrazolium Salt WST-8 as a Novel and Reliable Chromogenic Indicator for the Assessment of Boar Semen Quality.
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DOI：
10.3390/ani10122293
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发表时间：
2020-12-04
期刊：
影响因子：
--
通讯作者：
Jason Tsai PS
中科院分区：
文献类型：
--
作者：
Chen YH;Wu CP;Lin HL;Liaw RB;Lai YY;Wu MC;Chen LR;Jason Tsai PS
在人工授精中，良好的精液质量是繁殖成功的重要因素。然而，精液质量不能完全由精子活力或活力来评价。标准的精液质量评估需要昂贵的自动化计算机辅助精子分析或特定的和费力的标记程序，然后才能通过流式细胞术评估特定的参数。在本研究中，我们检查了广泛用于细胞生物学领域的2-[2-甲氧基-4-硝基苯基]-3-[4-硝基苯基]-5-[2，4-二磺基苯基]-2H-四唑（WST-8）测定法是否可用于评估精子活力，以及WST-8减少率是否与多个精子参数相关，这些参数与精子质量有关。我们在这项研究中证明，WST-8检测可以作为一种快速，经济，可靠的方法来预测精液质量的公猪。四唑鎓盐，2-[2-甲氧基-4-硝基苯基]-3-[4-硝基苯基]-5-[2，4-二磺基苯基]-2H-四唑鎓（WST-8），已被广泛用于测定细胞活力;然而，由于该测定仅提供关于细胞活力的信息，其在生殖领域中的应用仍然受到限制。本研究的目的是将WST-8减少率与各种精子质量相关参数（即，精子活力、运动力、前向运动力、顶体完整性和线粒体完整性），以便为公猪精液质量评价提供快速、可靠和可负担的评估。使用不同比例的活性/受损精子细胞，我们首先通过标准流式细胞术和计算机辅助精子分析来验证我们的样品制备。进一步分析表明，获得可靠预测模型的最有效实验条件是当精子浓度达到300 × 106个细胞/mL，精液/细胞计数试剂盒-8（CCK-8®）比例为200/10，孵育时间为20 min时。（光密度阅读值的差异，450 nm处的ΔOD）和精子参数高度相关（p < 0.01）。在精液样本有限的情况下，最低精液浓度为150 × 106个细胞/mL，精液/CCK-8®比例为200/20，孵育时间为30 min，仍可以使用WST-8测定法提供可靠的精子参数预测。我们的数据提供了强有力的证据，第一次，WST-8检测可用于评估公猪精液质量具有很大的潜力，可应用于不同的哺乳动物物种。
Good semen quality is an essential factor for breeding success when artificial insemination is performed. However, semen quality can not fully be evaluated by merely sperm viability or motility. Standard semen quality evaluation requires costly automated computer-assisted sperm analysis or specific and laborious labeling procedures before particular parameters can be assessed by flow cytometry. In the current study, we examined whether the 2-[2-methoxy-4-nitrophenyl]-3-[4-nitrophenyl]-5-[2,4-disulfophenyl]-2H-tetrazolium (WST-8) assay, which is widely used in the cell biology field, can be applied to evaluate sperm viability, and moreover, whether the WST-8 reduction rate can correlate with multiple sperm parameters that are related to sperm quality. We demonstrated in this study that the WST-8 assay can be used as a rapid, affordable, and reliable method for the prediction of semen quality in boar. A tetrazolium salt, 2-[2-methoxy-4-nitrophenyl]-3-[4-nitrophenyl]-5-[2,4-disulfophenyl]-2H-tetrazolium (WST-8), has been used widely to determine cell viability; however, its application in the field of reproduction is still limited due to this assay merely providing information regarding cell viability. The aim of this study was to correlate the WST-8 reduction rate with various sperm quality-related parameters (i.e., sperm viability, motility, progressive motility, acrosome integrity and mitochondria integrity) in order to provide a rapid, reliable and affordable assessment for boar semen quality evaluation. Using different ratios of active/damaged sperm cells, we first validated our sample preparations by standard flow cytometry and computer-assisted sperm analysis. Further analyses demonstrated that the most efficient experimental condition for obtaining a reliable prediction model was when sperm concentration reached 300 × 106 cells/mL with the semen/cell-counting kit-8 (CCK-8®) ratio of 200/10 and incubated time of 20 min. Under this set up, the WST-8 reduction rate (differences on optic density reading value, ΔOD at 450 nm) and sperm parameters were highly correlated (p < 0.01) for all sperm parameters evaluated. In the case of limited semen samples, a minimal semen concentration at 150 × 106 cells/mL with the semen/CCK-8® ratio of 200/20 and incubation time for 30 min could still provide reliable prediction of sperm parameters using the WST-8 assay. Our data provide strong evidence for the first time that the WST-8 assay could be used to evaluate boar semen quality with great potential to be applied to different mammalian species.
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DOI：
10.1016/s1387-2656(05)11004-7
发表时间：
2005-01-01
期刊：
BIOTECHNOLOGY ANNUAL REVIEW, VOL 11
影响因子：
--
作者：
Berridge, MV;Herst, PM;Tan, AS
通讯作者：
Tan, AS
影响因子：
1.6
作者：
Chang, Yu-Chia;Yu, Jane-Fang;Tsai, Pei-Shiue
通讯作者：
Tsai, Pei-Shiue
影响因子：
5.5
作者：
Kairo, SK;Bedwell, J;Corbel, MJ
通讯作者：
Corbel, MJ
DOI：
10.1007/978-1-61779-108-6_4
发表时间：
2011-01-01
期刊：
MAMMALIAN CELL VIABILITY: METHODS AND PROTOCOLS
影响因子：
--
作者：
Stoddart, Martin J.
通讯作者：
Stoddart, Martin J.
影响因子：
3.5
作者：
Bucher, K.;Malama, E.;Bollwein, H.
通讯作者：
Bollwein, H.