Department of Theriogenology, University of Veterinary and Animal Sciences, Lahore 54000, Pakistan.
J Anim Sci. 2010 Mar;88(3):922-5. doi: 10.2527/jas.2009-2344. Epub 2009 Dec 18.
The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, which measures the reduction of MTT, is commonly used to validate the viability of metabolically active cells. This study was conducted to evaluate and validate the MTT assay to assess the spermatozoal viability of Nili-Ravi buffalo bulls and compare the efficiency of the test with the supravital staining technique (eosin and nigrosin) and the hypoosmotic swelling test. Fresh semen samples from breeding Nili-Ravi buffalo bulls (n = 25) were collected using an artificial vagina. After assessing the quality of the semen for routine variables, the MTT assay was carried out in PBS. Results revealed a correlation (r = 0.979; P < 0.001) between the viability of spermatozoa and the rate of reduction of MTT. The other proportions of same semen samples showed a poor relationship between the eosin and nigrosin test (r = -0.25), the hypoosmotic swelling test (r = -0.12), and motility (r = -0.15). However, the MTT assay was found to be superior compared with other tests because it was able to determine those spermatozoa that were more than 90% viable. In conclusion, the MTT assay is a simple, robust test that can be used to select Nili-Ravi buffalo bulls on the basis of spermatozoa quality.
3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)测定法，用于测量 MTT 的还原，常用于验证代谢活跃细胞的活力。本研究旨在评估和验证 MTT 测定法，以评估尼里-拉菲水牛公牛精子的活力，并将该测试的效率与台盼蓝染色技术(伊红和黑固红)和低渗肿胀试验进行比较。使用人工授精阴道采集了 25 头繁殖尼里-拉菲水牛公牛的新鲜精液样本。在评估常规变量精液质量后，在 PBS 中进行 MTT 测定。结果显示，精子活力与 MTT 还原率之间存在相关性(r = 0.979; P < 0.001)。同一精液样本的其他比例显示，伊红和黑固红试验(r = -0.25)、低渗肿胀试验(r = -0.12)和运动性(r = -0.15)之间的相关性较差。然而，与其他测试相比，MTT 测定法被发现具有优势，因为它能够确定超过 90%活力的精子。总之，MTT 测定法是一种简单、稳健的测试方法，可用于根据精子质量选择尼里-拉菲水牛公牛。
