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Sperm MTT viability assay: a new method for evaluation of human sperm viability.精子MTT活力测定：一种评估人类精子活力的新方法。
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Suitability of the hypo-osmotic swelling test for assessing the viability of cryopreserved sperm.低渗肿胀试验用于评估冷冻保存精子活力的适用性。
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Establishment of an optimal hypo-osmotic swelling test by examining single spermatozoa in four different hypo-osmotic solutions.通过在四种不同的低渗溶液中检测单个精子来建立最佳低渗肿胀试验。
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Comparison of different hypo-osmotic swelling solutions to select viable immotile spermatozoa for potential use in intracytoplasmic sperm injection.比较不同低渗肿胀溶液以选择可用于胞浆内单精子注射的有活力的不动精子。
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Microscopic analysis of MTT stained boar sperm cells.对经MTT染色的公猪精子细胞进行显微镜分析。
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Selection of Sperm Based on Hypo-Osmotic Swelling May Improve ICSI Outcome: A Preliminary Prospective Clinical Trial.基于低渗肿胀法选择精子可能改善卵胞浆内单精子注射结局：一项初步前瞻性临床试验
Int J Fertil Steril. 2014 Apr;8(1):21-8. Epub 2014 Mar 9.

本文引用的文献

1
The use of a modified hypo-osmotic swelling test for the selection of viable ejaculated and testicular immotile spermatozoa in ICSI.在卵胞浆内单精子注射中使用改良低渗肿胀试验筛选有活力的射出及睾丸来源的不动精子。
Hum Reprod. 2001 Feb;16(2):272-6. doi: 10.1093/humrep/16.2.272.
2
Developmental capacity of damaged spermatozoa.受损精子的发育能力。
Hum Reprod. 1999 Sep;14(9):2279-85. doi: 10.1093/humrep/14.9.2279.
3
Time course of hypo-osmotic swellings of human spermatozoa: evidence of ordered transition between swelling subtypes.人类精子低渗肿胀的时间进程：肿胀亚型之间有序转变的证据。
Hum Reprod. 1998 Jun;13(6):1578-83. doi: 10.1093/humrep/13.6.1578.
4
Effect of pentoxifylline on immotile testicular spermatozoa.己酮可可碱对不动的睾丸精子的作用。
J Assist Reprod Genet. 1998 Feb;15(2):90-2. doi: 10.1007/BF02766832.
5
High fertilization rate obtained after intracytoplasmic sperm injection with 100% nonmotile spermatozoa selected by using a simple modified hypo-osmotic swelling test.通过使用简单改良低渗肿胀试验选择的100%无运动能力精子进行胞浆内单精子注射后获得了高受精率。
Fertil Steril. 1997 Aug;68(2):373-5. doi: 10.1016/s0015-0282(97)81533-6.
6
The single sperm curling test, a modified hypo-osmotic swelling test, as a potential technique for the selection of viable sperm for intracytoplasmic sperm injection.单精子卷曲试验，一种改良的低渗肿胀试验，作为一种为胞浆内单精子注射选择活精子的潜在技术。
Fertil Steril. 1997 Aug;68(2):346-50. doi: 10.1016/s0015-0282(97)81527-0.
7
Establishment of an optimal hypo-osmotic swelling test by examining single spermatozoa in four different hypo-osmotic solutions.通过在四种不同的低渗溶液中检测单个精子来建立最佳低渗肿胀试验。
Hum Reprod. 1997 May;12(5):1111-3. doi: 10.1093/humrep/12.5.1111.
8
The hypo-osmotic swelling test for selection of viable sperm for intracytoplasmic sperm injection in men with complete asthenozoospermia.用于为严重少弱精子症男性选择可行精子进行卵胞浆内单精子注射的低渗肿胀试验。
Fertil Steril. 1996 May;65(5):972-6. doi: 10.1016/s0015-0282(16)58271-5.
9
Development of an assay to assess the functional integrity of the human sperm membrane and its relationship to other semen characteristics.一种用于评估人类精子膜功能完整性及其与其他精液特征关系的检测方法的开发。
J Reprod Fertil. 1984 Jan;70(1):219-28. doi: 10.1530/jrf.0.0700219.
10
Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays.用于细胞生长和存活的快速比色测定法：应用于增殖和细胞毒性测定。
J Immunol Methods. 1983 Dec 16;65(1-2):55-63. doi: 10.1016/0022-1759(83)90303-4.

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.

DOI:10.1023/a:1020310503143
PMID:12416652
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3455683/
Abstract

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.

摘要

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。