https://scholars.lib.ntu.edu.tw/handle/123456789/573176
標題: | Tetrazolium salt WST-8 as a novel and reliable chromogenic indicator for the assessment of boar semen quality | 作者: | Chen Y.-H Wu C.-P Lin H.-L Liaw R.-B Lai Y.-Y Wu M.-C Chen L.-R Tsai P.-S.J. PEI-SHIUE TSAI |
關鍵字: | chromogenic substrate; reactive oxygen metabolite; tetrazolium; acrosome; apoptosis; Article; cell viability; cell viability assay; controlled study; cryopreservation; dairy cattle; flow cytometry; human; human cell; mitochondrial membrane potential; mitochondrion; optical density; pig; semen analysis; sperm quality; spermatozoon; spermatozoon density; spermatozoon motility; WST-8 assay | 公開日期: | 2020 | 卷: | 10 | 期: | 12 | 起(迄)頁: | 1-12 | 來源出版物: | Animals | 摘要: | A tetrazolium salt, 2-[2-methoxy-4-nitrophenyl]-3-[4-nitrophenyl]-5-[2,4-disulfophenyl]-2Htetrazolium (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, ?OD 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. ? 2020 by the authors. Licensee MDPI, Basel, Switzerland. |
URI: | https://www.scopus.com/inward/record.uri?eid=2-s2.0-85097311775&doi=10.3390%2fani10122293&partnerID=40&md5=026cd21bb13c1b81f2325ef144fbf963 https://scholars.lib.ntu.edu.tw/handle/123456789/573176 |
ISSN: | 20762615 | DOI: | 10.3390/ani10122293 |
顯示於: | 獸醫學系 |
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