精子MTT活力测定：一种评估人类精子活力的新方法。
Sperm MTT viability assay: a new method for evaluation of human sperm viability.
作者信息
Nasr-Esfahani Mohammad Hossein, Aboutorabi Roshanak, Esfandiari Ebrahim, Mardani Mohammad
机构信息
Department of Clinical and Experimental Embryology and Andrology, Royan Institute, Tehran, Iran.
出版信息
J Assist Reprod Genet. 2002 Oct;19(10):477-82. doi: 10.1023/a:1020310503143.
PURPOSE
MTT (3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay is commonly used as a cell proliferation assay. The objective of this study was to evaluate the capability of MTT assay to discriminate between viable and nonviable sperms and compare it sefficiency with E&N (eosin and nigrosin) and HOST (hypo-osmotic swelling test).
METHODS
MTT assay was modified to obtain optimal result for assessment of sperm viability. After standardization of method, MTT, E&N, and HOST were carried out simultaneously on 57 semen samples from patients referring to Isfahan Fertility and Infertility Center. The correlation coefficient between these tests and sperm motility was calculated using the SPSS statistical program. Specificity and sensitivity of each test was also obtained.
RESULTS
The optimal conditions for sperm MTT viability assay were 2 h after addition of sperm to MTT in HAM'S F10 + 25 mM HEPES + 10% HSA at 37 degrees C and pH 7.4-7.45. Inter- and intra-assay coefficients of variations were 9 and 7%, respectively. The sensitivity and specificity for sperm MTT viability assay, E&N, and HOST were 97,98, and 99%, and 100, 100, and 83% respectively. High significant correlations were obtained between sperm MTT viability assay, E&N, HOST and motility.
CONCLUSIONS
Sperm MTT viability assay can be used as a diagnostic test for discrimination of viable sperms from sperm population.
目的
MTT（3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐）检测常用于细胞增殖检测。本研究的目的是评估MTT检测区分活精子与死精子的能力，并将其效率与伊红-苯胺黑（E&N）染色法和低渗肿胀试验（HOST）进行比较。
方法
对MTT检测进行改良以获得评估精子活力的最佳结果。方法标准化后，对转诊至伊斯法罕生育与不孕中心的57例患者的精液样本同时进行MTT检测、E&N染色法和HOST检测。使用SPSS统计软件计算这些检测方法与精子活力之间的相关系数。还获得了每种检测方法的特异性和敏感性。
结果
精子MTT活力检测的最佳条件为：在含25 mM HEPES、10%人血清白蛋白（HSA）的哈姆氏F10培养基中，于37℃、pH 7.4 - 7.45条件下加入精子后2小时进行MTT检测。批间和批内变异系数分别为9%和7%。精子MTT活力检测、E&N染色法和HOST检测的敏感性分别为97%、98%和99%，特异性分别为100%、100%和83%。精子MTT活力检测、E&N染色法、HOST检测与精子活力之间均存在高度显著相关性。
结论
精子MTT活力检测可作为一种诊断方法，用于从精子群体中区分活精子。
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