Smikle C B, Turek P J
Reproductive Endocrinology Center, University of California School of Medicine, San Francisco, USA.
Mol Reprod Dev. 1997 Jun;47(2):200-3. doi: 10.1002/(SICI)1098-2795(199706)47:2<200::AID-MRD11>3.0.CO;2-3.
Viable, healthy sperm are preferred for oocyte fertilization with intracytoplasmic sperm injection. Currently, motility is the most widely applied measure of sperm viability. However, with this criterion, viable but immotile sperm are overlooked as candidates for micromanipulation. Supravital stains identify viable sperm but may be toxic to the gamete or embryo. We tested the hypothesis that hypo-osmotic swelling, developed to assess sperm membrane integrity, can accurately determine sperm viability. Thawed sperm from 12 fertile donors were exposed to a hypo-osmotic solution and to two supravital stains. A total of 2,010 sperm were assessed for tail coiling after a 30-min exposure to hypoosmotic solution. By supravital stains, 31% of thawed sperm were viable; 23% showed tail coiling. Among coiled-tail sperm, 100% were viable by supravital stain. As a measure of viability, tail coiling exhibited a sensitivity of 30%, specificity of 100%, and positive predictive value of 100% compared to supravital stains. With a 60-min hypo-osmotic incubation, the specificity (89%) and positive predictive value (78%) decreased significantly (P < 0.05). Therefore, hypo-osmotic swelling accurately detects viable sperm among a nonmotile population. Assay accuracy, however, is very sensitive to the incubation time in hypo-osmotic solution.
卵胞浆内单精子注射技术进行卵母细胞受精时，优选有活力的健康精子。目前，运动能力是评估精子活力应用最广泛的指标。然而，基于这一标准，有活力但不运动的精子作为显微操作的候选对象被忽视了。体外活体染色可识别有活力的精子，但可能对配子或胚胎有毒性。我们检验了这样一个假设：用于评估精子膜完整性的低渗肿胀试验能够准确测定精子活力。将12名生育能力正常的供体解冻后的精子分别置于低渗溶液和两种体外活体染液中。在低渗溶液中孵育30分钟后，共对2010条精子的尾部卷曲情况进行了评估。通过体外活体染色发现，解冻后的精子中有31%具有活力；23%出现尾部卷曲。在尾部卷曲的精子中，通过体外活体染色显示100%具有活力。作为一种活力检测方法，与体外活体染色相比，尾部卷曲试验的灵敏度为30%，特异性为100%，阳性预测值为100%。低渗孵育60分钟时，特异性（89%）和阳性预测值（78%）显著降低（P<0.05）。因此，低渗肿胀试验能准确检测出不活动群体中的有活力精子。然而，检测准确性对低渗溶液中的孵育时间非常敏感。
